| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NBP2-93116-0.02ml | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NBP2-98113-50ul | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NBP1-21502SS | |||
| Conjugate | Catalog # | Availability | Size | Price |
|---|---|---|---|---|
| Alexa Fluor 405 | NB100-182AF405 | |||
| Alexa Fluor 488 | NB100-182AF488 | |||
| Alexa Fluor 647 | NB100-182AF647 | |||
| Alexa Fluor 700 | NB100-182AF700 | |||
| Biotin | NB100-182B | |||
| DyLight 405 | NB100-182V | |||
| DyLight 488 | NB100-182G | |||
| DyLight 550 | NB100-182R | |||
| DyLight 650 | NB100-182C | |||
| FITC | NB100-182F | |||
| HRP | NB100-182H | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NB100-182SS | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NBP2-22164SS | |||
| Azide and BSA Free | NBP2-80954 | |||
![Immunohistochemistry-Paraffin: SLC3A1 Antibody [NBP2-47272] - Staining in human kidney and cervix, uterine tissues using anti-SLC3A1 antibody. Corresponding SLC3A1 RNA-seq data are presented for the same tissues.](http://images.novusbio.com/fullsize/SLC3A1-Antibody-Immunohistochemistry-Paraffin-NBP2-47272-img0004.jpg "<b>Orthogonal Strategies Validation. </b>Immunohistochemistry-Paraffin: SLC3A1 Antibody [NBP2-47272] - Staining in human kidney and cervix, uterine tissues using anti-SLC3A1 antibody. Corresponding SLC3A1 RNA-seq data are presented for the same tissues.")
![<b>Orthogonal Strategies Validation. </b>Western Blot: SLC3A1 Antibody [NBP2-47272] - Analysis in human small intestine tissue.](http://images.novusbio.com/fullsize/SLC3A1-Antibody-Western-Blot-NBP2-47272-img0005.jpg)
![Immunohistochemistry-Paraffin: SLC3A1 Antibody [NBP2-47272] - Staining of human cervix, uterine shows low expression as expected.](http://images.novusbio.com/fullsize/SLC3A1-Antibody-Immunohistochemistry-Paraffin-NBP2-47272-img0003.jpg)
![Immunohistochemistry-Paraffin: SLC3A1 Antibody [NBP2-47272] - Staining of human kidney shows strong cytoplasmic and membrane positivity in cells in tubules.](http://images.novusbio.com/fullsize/SLC3A1-Antibody-Immunohistochemistry-NBP2-47272-img0001.jpg)
![Western Blot: SLC3A1 Antibody [NBP2-93116] - Analysis of extracts of various cell lines, using SLC3A1 at 1:1000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit . Exposure time: 90s.](http://images.novusbio.com/fullsize/SLC3A1-Antibody-Western-Blot-NBP2-93116-img0002.jpg "Western Blot: SLC3A1 Antibody [NBP2-93116] - Analysis of extracts of various cell lines, using SLC3A1 at 1:1000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit . Exposure time: 90s.")
![Immunohistochemistry-Paraffin: SLC3A1 Antibody [NBP2-93116] - Mouse kidney using SLC3A1 Rabbit pAb at dilution of 1:200 (40x lens).](http://images.novusbio.com/fullsize/SLC3A1-Antibody-Immunohistochemistry-Paraffin-NBP2-93116-img0004.jpg)
![Immunohistochemistry-Paraffin: SLC3A1 Antibody [NBP2-93116] - Rat kidney using SLC3A1 Rabbit pAb at dilution of 1:200 (40x lens).](http://images.novusbio.com/fullsize/SLC3A1-Antibody-Immunohistochemistry-Paraffin-NBP2-93116-img0003.jpg)
![Western Blot: SLC3A1 Antibody [NBP2-68912] - Analysis in human cell line RT-4, human cell line U-251 MG, human plasma, human liver tissue and human tonsil tissue.](http://images.novusbio.com/fullsize/SLC3A1-Antibody-Western-Blot-NBP2-68912-img0001.jpg "Western Blot: SLC3A1 Antibody [NBP2-68912] - Analysis in human cell line RT-4, human cell line U-251 MG, human plasma, human liver tissue and human tonsil tissue.")
![Immunocytochemistry/Immunofluorescence: SLC3A1 Antibody [NBP2-68912] - Staining of human cell line RPTEC TERT1 shows localization to mitochondria.](http://images.novusbio.com/fullsize/SLC3A1-Antibody-Immunocytochemistry-Immunofluorescence-NBP2-68912-img0002.jpg)
![Western Blot: SLC3A1 Antibody [NBP2-85757] - Host: Rabbit. Target Name: SLC3A1. Sample Tissue: Human Hela Whole Cell lysates. Antibody Dilution: 1ug/ml](http://images.novusbio.com/fullsize/SLC3A1-Antibody-Western-Blot-NBP2-85757-img0001.jpg "Western Blot: SLC3A1 Antibody [NBP2-85757] - Host: Rabbit. Target Name: SLC3A1. Sample Tissue: Human Hela Whole Cell lysates. Antibody Dilution: 1ug/ml")
![Immunocytochemistry/Immunofluorescence: SLC3A1 Antibody [NBP2-98113] - Immunofluorescence staining of SLC3A1 in HeLa cells. Cells were fixed with 4% PFA, permeabilzed with 0.1% Triton X-100 in PBS, blocked with 10% serum, and incubated with NBP2-98113 antibody (dilution ratio 1:200) at 4C overnight. Then cells were stained with the Alexa Fluor(R)488-conjugated Goat Anti-rabbit IgG secondary antibody (green) and counterstained with DAPI (blue).Positive staining was localized to Cytoplasm.](http://images.novusbio.com/fullsize/nbp2-98113_rabbit-polyclonal-slc3a1-antibody-2352025112939.png "Immunocytochemistry/Immunofluorescence: SLC3A1 Antibody [NBP2-98113] - Immunofluorescence staining of SLC3A1 in HeLa cells. Cells were fixed with 4% PFA, permeabilzed with 0.1% Triton X-100 in PBS, blocked with 10% serum, and incubated with NBP2-98113 antibody (dilution ratio 1:200) at 4C overnight. Then cells were stained with the Alexa Fluor(R)488-conjugated Goat Anti-rabbit IgG secondary antibody (green) and counterstained with DAPI (blue).Positive staining was localized to Cytoplasm.")
![HepG2 cells were fixed and permeabilized for 10 minutes using -20C MeOH. The cells were incubated with Ferroportin/SLC40A1 Antibody at 5.0 ug/ml overnight at 4C and detected with an anti-rabbit DyLight 488 (green) at a 1:500 dilution. Alpha tubulin (DM1A) [NB100-690] was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (red) at a 1:500 dilution. Nuclei were counterstained with DAPI (blue). Cells were imaged using a 40X objective.](http://images.novusbio.com/fullsize/Ferroportin-SLC40A1-Antibody---BSA-Free-Immunocytochemistry-Immunofluorescence-NBP1-21502-img0012.jpg)
![Immunohistochemistry-Paraffin: Rabbit Polyclonal Ferroportin/SLC40A1 Antibody [NBP1-21502] - Staining of mouse small intestines using Ferroportin/SLC40A1 Antibody. Tissue was fixed in formalin overnight and embedded in paraffin. ~5 micro sections on slides were dehydrated. Antigen retrieval was done using citrate buffer in rice steamer for 20mins. 1 hour blocking without permeabilization. Overnight primary ab incubation was done 1:200 (NBP1-21502) at 4C. 2-hr secondary antibody incubation was at room temp; mounting media with DAPI. Image from a verified customer review.](http://images.novusbio.com/fullsize/antibody/nbp1-21502_rabbit-polyclonal-ferroportin-slc40a1-antibody-immunohistochemistry-paraffin-165202493537..jpg)
![Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Analysis of Cu & Fe content & metal-binding protein expression in wild-type & PrPC-null mouse total brain at different ages. (A) The graph shows the ratio of Cu & Fe levels in Prnp0/0 & Prnp+/+ brain samples (P1, P365 N = 4; P7, P30, P180 N = 6; P90 N = 5). (B) Representative Western blot images showing metal-binding protein levels in Prnp0/0 & Prnp+/+ brain samples (N = 4). The constant level of the housekeeping proteins ( beta -III Tubulin & beta -Actin) are also reported. (C) The graph shows the up- or down-regulation of protein expression in Prnp0/0 samples compared to Prnp+/+, i.e., (Prnp0/0 protein OD/housekeeping OD)/(Prnp+/+ protein OD/housekeeping OD). All error bars indicate SD; N = 4 minimum; *p 0.05 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28864822), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-21502_rabbit-polyclonal-ferroportin-slc40a1-antibody-310202415382421.jpg)
![Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - SHP abrogates the BMP6 effect on iron metabolism through inhibition of hepcidin gene expression in mice.(a–d) C57/BL6 mice were injected with Ad-GFP (n = 4 per group, 5.9 × 109 pfu) or Ad-Flag-SHP (n = 5 per group, 5.9 × 109 pfu) via the tail-vein, & treated with Vehicle or BMP6 (500 μg/kg, i.p.) for 6 h at day 5 after the infection. (a) Serum iron level. (b) Q-PCR analysis showing hepcidin & SHP mRNA levels in liver. (c) Serum hepcidin levels. (d) Western blot analysis showing SMAD1/5/8 phosphorylation & SHP expression in liver. (e) Western blot analysis showing FPN expression in spleen (top). Graphical representation showing FPN expression (bottom). Data are presented as means ± SD. Arrows show locations of molecular weight markers. The western blot images were cropped with a grey cropping line. All gels for western blot analysis were run under the same experimental conditions. **P < 0.01, ***P < 0.001 by two-tailed Student t-test. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep34630), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-21502_rabbit-polyclonal-ferroportin-slc40a1-antibody-310202416165613.jpg)
![Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Upregulation of H3K4me3 increases iron exporter Fpn1 & exhibits neuroprotection.a, b CPI-455 treatment (24 h) increased the protein level of H3K4me3 & Fpn1 in SH-SY5Y cells. c MTT assay showed CPI-455 showed neuroprotection against the toxicity of 6-OHDA in SH-SY5Y cells. d–g SH-SY5Y cells were infected by AAV-KDM5C-silencing virus, & KDM5C was significantly decreased at the mRNA level as reflected by RT-PCR d & at the protein level by Western blot e, f. In addition, Fpn1 & H3K4me3 were increased in AAV-KDM5C-silencing virus infected cells e, f. Moreover, g silencing KDM5C showed a slight neuroprotection against 6-OHDA-induced PD model in SH-SY5Y cells. *P < 0.05; **P < 0.01; ***P < 0.001. Data are presented as mean ± SEM. n = 3–4 for each group in Western blot & n = 5 for each group in MTT assay. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33116116), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-21502_rabbit-polyclonal-ferroportin-slc40a1-antibody-31020241616071.jpg)
![Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Upregulation of H3K4me3 increases iron exporter Fpn1 & exhibits neuroprotection.a, b CPI-455 treatment (24 h) increased the protein level of H3K4me3 & Fpn1 in SH-SY5Y cells. c MTT assay showed CPI-455 showed neuroprotection against the toxicity of 6-OHDA in SH-SY5Y cells. d–g SH-SY5Y cells were infected by AAV-KDM5C-silencing virus, & KDM5C was significantly decreased at the mRNA level as reflected by RT-PCR d & at the protein level by Western blot e, f. In addition, Fpn1 & H3K4me3 were increased in AAV-KDM5C-silencing virus infected cells e, f. Moreover, g silencing KDM5C showed a slight neuroprotection against 6-OHDA-induced PD model in SH-SY5Y cells. *P < 0.05; **P < 0.01; ***P < 0.001. Data are presented as mean ± SEM. n = 3–4 for each group in Western blot & n = 5 for each group in MTT assay. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33116116), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-21502_rabbit-polyclonal-ferroportin-slc40a1-antibody-31020241616370.jpg)
![Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - SHP deficiency alters hepcidin gene expression in liver of HID mice.(a–i) WT & SHP KO mice (n = 5 per group) were fed with high-iron diet (HID, 8 g/kg) for 3 weeks. (a) Serum iron level. (b) Hepcidin mRNA level in liver. (c) Hepcidin expression in mouse liver. IHC was performed using an antibody against hepcidin. Scale bar shows 50 μm. (d) Serum hepcidin level. (e) Western blot analysis (left panel) showing hepatic SHP & splenic FPN expression & graphical representation (right panel) showing splenic FPN expression. (f) Splenic iron level. (g) Perls’ prussian blue staining in spleen. Scale bar shows 200 μm. (h) BMP6 & BMP9 mRNA levels in liver. (i) Western blot analysis (top) & graphical representation (bottom) showing SMAD1/5/8 phosphorylation in liver. The grouping of the images is from different parts of the same gel. Data are presented as means ± SD. Arrows show locations of molecular weight markers. The experiment was repeated on a minimum of three separate occasions. The western blot images were cropped with a grey cropping line. All gels for western blot analysis were run under the same experimental conditions. *P < 0.05, **P < 0.01, ***P < 0.001 by two-tailed Student t-test. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep34630), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-21502_rabbit-polyclonal-ferroportin-slc40a1-antibody-310202416173542.jpg)
![Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Differential expression of proteins of iron metabolism in CCA cells. The CCA4, CCLP1 & HUCCT1 human CCA-derived cell lines were cultured as adherent monolayers (MON) or in 3D sphere conditions (SPH). Panel a. Top left, representative immunoblot analysis. Cell extracts were reacted with antibodies against transferrin receptor (TfR1), ferroportin (FPN), ferritin H subunit (FtH) & vinculin. Cropped blots are displayed. The original full blot images can be found in Supplementary Information. The graphs show densitometric quantification of immunoblot analyses. The values were normalized to vinculin & expressed as a fraction of respective MON cells normalized to 1. Mean values ± SEM (n = 6), *p ≤ 0.05, **p ≤ 0.01 vs control MON for each cell line. Panel b. RNA bandshift analysis of IRP activity. Cytoplasmic extracts were incubated with a 32P-labeled iron-responsive element (IRE) probe & RNA-protein complexes separated on non-denaturing polyacrylamide gels. On the left a representative autoradiogram is shown. A cropped gel is displayed. The original full gel image can be found in Supplementary Information. The graph on the right shows the densitometric quantification of IRPs bands by direct nuclear counting, as described in Materials & Methods; mean percentages ± SEM of control values (n = 6),, **p ≤ 0.01, ***p< 0.001 vs control MON for each cell line. Panel c. TfR1 & FPN mRNA levels were measured by quantitative RT-PCR. Samples were analyzed in triplicate, normalized to the housekeeping gene 18 S & expressed as percentage of respective MON cells normalized to 1. Mean values ± SEM (n = 6), **p ≤ 0.01 vs control MON for each cell line. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29247214), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-21502_rabbit-polyclonal-ferroportin-slc40a1-antibody-31020241616561.jpg)
![Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Iron (55Fe) transport across the apical & basal borders. (a) Apical transport (uptake) of 55Fe in response to intracellular/basal ascorbate (150 μM) was increased compared to no ascorbate. The cells were incubated with ascorbate in the basal chamber for 24 h. Values are means of 10 samples ±SD. The difference between treatments was significant (p = 0.04). (b) Basolateral transport of iron (as 55Fe) in response to intracellular/basal ascorbate (150 μM) was increased compared to no ascorbate. The cells were incubated with ascorbate in the basal chamber for 24 h. Values are means of 10 samples ±SD. The difference between treatments was significant (p = 0.03). (c) Western Blot of cells treated with ascorbate (150 μM). Lane 1: Control cells (no treatment). Lane 2: Basal ascorbate at 150 μM; these were the ferroportin levels before iron addition in (a). Lane 3: Ferroportin levels 22 h after the iron addition (20 μM for 2 h) in (a). Image collected & cropped by CiteAb from the following publication (//www.mdpi.com/2072-6643/6/1/249), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-21502_rabbit-polyclonal-ferroportin-slc40a1-antibody-310202416171416.jpg)
![Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Metformin rescues BMP6-mediated alteration of iron metabolism in mice.(a–e) C57/BL6 mice (n = 4 per group) were treated with BMP6 (500 μg/kg, i.p.) & metformin (200 mg/kg, p.o.). (a) Serum iron level. (b) Q-PCR analysis showing hepcidin & SHP mRNA levels in liver. (c) Serum hepcidin levels. (d) Western blot analysis showing SMAD1/5/8 phosphorylation & FPN in liver. (e) Western blot analysis (top) & graphical representation (bottom) showing FPN expression in spleen. (f) Schematic diagram of SHP-mediated inhibition of BMP6-SMADs pathway. Data are presented as means ± SD. Arrows show locations of molecular weight markers. The western blot images were cropped with a grey cropping line. All gels for western blot analysis were run under the same experimental conditions. *P < 0.05, **P < 0.01, ***P < 0.001 by two-tailed Student t-test. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep34630), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-21502_rabbit-polyclonal-ferroportin-slc40a1-antibody-31020241616098.jpg)
![Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Comparison of Cu, Fe, & metal-binding protein expression levels in wild-type & PrPC-null mouse spleen at different ages. (A) The graph shows the ratio of Cu & Fe levels in Prnp0/0 & Prnp+/+ spleen samples (P15 N = 3; P30 N = 4; P90, P180 N = 6; P365 N = 5). (B) The graph shows the weight of spleen extracted from Prnp0/0 & Prnp+/+ mice; N = 4. (C) Representative Western blot images showing metal-binding protein levels in Prnp0/0 & Prnp+/+ spleen samples. The constant level of the housekeeping protein ( beta -Actin) are also reported. (D) The graph shows the up- or down-regulation of protein expression in Prnp0/0 samples compared to Prnp+/+, i.e., (Prnp0/0 protein OD/housekeeping OD)/ (Prnp+/+ protein OD/housekeeping OD); N = 4. All error bars indicate SD; *p < 0.05; **p < 0.01; ***p < 0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27729845), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-21502_rabbit-polyclonal-ferroportin-slc40a1-antibody-310202416173510.jpg)
![Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Htt elimination disrupts brain iron homeostasis.(A) Representative photographs of immunohistochemical staining for ferritin light chain (Ft). Left panels: cerebella of 21mo CTL noTM & 18mo cKO TM@3mo; middle panels: thalami of 12mo CTL TM@9mo & 12mo cKO TM@9mo; right panels: striata of 21mo CTL noTM & 18mo cKO TM@3mo. Note that Ft levels are already extremely reduced in the thalamus 3 months after Htt elimination (middle panels), & that Ft expression is practically abolished in the cerebellum in the absence of Htt (left panels). (B) Representative western blots of Tfr & Fpn protein expression in 10 month-old brains from CTL & cKO mice TM-treated at 6 months of age. Antibody against actin was used as internal control for loading. Note that Tfr & Fpn levels are increased in TM-treated cKO compared to controls. (C) Quantification of Tfr & Fpn expression levels. Western blots of total protein extracts from 10 month-old brains CTL (n = 5) & cKO (n = 5) TM-treated at 6 months of age were probed with anti-Tfr or anti-Fpn, stripped & re-probed with anti-actin antibody. Bands intensities were quantitated using Image J. Tfr & Fpn levels were normalized over actin levels. Values represent mean relative to controls ± SD (***P<0.001, Student’s t-test). Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pgen.1006846), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-21502_rabbit-polyclonal-ferroportin-slc40a1-antibody-310202416171422.jpg)
![Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - GW501516 activation of PPARδ regulates 6-OHDA-triggered expression of level of DMT1 & FPN1 protein, & DMT1 mRNA, but not FPN1 mRNA. (A) Cells were treated with 6-OHDA for the indicated durations. (B,C) Cells pretreated with DMSO or GW501516 for 8 h were incubated with or without 6-ODHA for 16 h. (D) Cells were treated with GW501516 for the indicated durations. (E,F) Cells pretreated with DMSO or GW501516 for 8 h were incubated with or without 6-ODHA for 16 h. Total RNA & protein were extracted, & mRNA & protein levels were analyzed by real-time PCR (A,B,D,E) & Western blot (C,F), respectively. RPS18 & alpha -tubulin were used as internal controls for real-time PCR & Western blot, respectively. Results are expressed as means ± SE (n = 3). An image analyzer was used to quantify band intensity of Western blot, & the ratio of protein to alpha -tubulin is indicated above each lane. * p < 0.05, ** p < 0.01 relative to the untreated group; #p < 0.05, ##p < 0.01 relative to the 6-OHDA-treated group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35624674), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-21502_rabbit-polyclonal-ferroportin-slc40a1-antibody-310202416171453.jpg)
![Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Fisetin protects against DOX-induced ferroptosis via regulating Nrf2 in rats. (A). Western blot results of Nrf2 & Keap1, HO-1, FTH1, FTL, FPN, & TfR1 protein in control & different drug-treated rat heart tissues. (B). Quantification of (A). (C). Expression of Nrf2 was detected by immunohistochemistry (IHC) (Representative images, 200X & 400X, Scale bar = 100 & 50 μm, n = 6 rats per groups) in cardiac tissue of each group. The values are presented as mean ± SD. *p < 0.05; **p < 0.01; ***p < 0.001; NS, no significance; DOX, doxorubicin. Nrf2, nuclear factor erythroid 2-related factor 2; Keap1, Kelch-like ECH-associated protein 1; HO-1, heme oxygenase-1; FTH1, ferritin heavy chain 1; FTL, ferritin light chain; FPN, ferroportin; TfR1, transferrin receptor 1. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35273493), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-21502_rabbit-polyclonal-ferroportin-slc40a1-antibody-31020241616065.jpg)
![Immunocytochemistry/ Immunofluorescence: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Distribution of TfR & Fpn in the anterior segment of bovine eye:(A) The reaction for TfR is evident on the ciliary epithelium facing the AH (panel 1). TfR is localized on the apical membrane & intracellular vesicles of NPE cells as in the human sample (panel 2). The corneal epithelium does not react for TfR (panel 3), while the corneal endothelium shows a distinct reaction on the plasma membrane (panel 4). H&E staining shows multiple layers of corneal epithelium, the stroma, & a single layer of corneal endothelium (panel 5). Scale bar: 25 μm. (B) Fpn is expressed on the ciliary epithelium & the iris (panel 1). The expression of Fpn is limited to NPE cells as in the human sample (panel 2). H&E staining shows the orientation of ciliary epithelium with respect to the iris & AH (panel 3). Unlike human samples, the bovine corneal epithelium did not react for Fpn (panel 4). However, corneal endothelium showed strong reactivity for Fpn (panel 5). Scale bar: 25 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29859760), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-21502_rabbit-polyclonal-ferroportin-slc40a1-antibody-31020241618269.jpg)
![Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - GW501516 PPARδ activation suppresses 6-OHDA-induced expression of IRP1 in SH-SY5Y cells. (A,B) Cells were treated with 6-OHDA (A) or GW501516 (B) for the indicated durations. (C) Cells stably expressing shRNA targeting scrambled sequences or PPARδ were pretreated with DMSO or GW501516 for 8 h & subsequently incubated in the presence or absence of 6-ODHA for 16 h. (D,E) Cells pretreated with DMSO or GW501516 for 8 h were incubated with or without 6-ODHA for 16 h. Total RNA & protein were extracted, & levels of mRNA & protein were analyzed by real-time PCR (A–D) & Western blot (E). The immunoblots are separate from those shown in Figure 4C,F. RPS18 & alpha -tubulin were used as internal controls for real-time PCR & Western blot, respectively. Results are expressed as means of triplicate ± SE (A–D). * p < 0.05, ** p < 0.01 relative to untreated group; #p < 0.05 relative to 6-OHDA-treated group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35624674), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-21502_rabbit-polyclonal-ferroportin-slc40a1-antibody-3102024161602.jpg)
![Immunocytochemistry/ Immunofluorescence: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Expression of iron modulating proteins in the cornea & ciliary body.(A) Probing of lysates from bovine cornea (Cor) & CB for Cp, TfR, Tf, & ferritin shows the expression of all of these proteins in both samples (lanes 1–4). Deglycosylation results in faster migration of Cp, TfR, & Tf on SDS-PAGE, indicating the presence of glycans (lanes 2 & 4). AH & the vitreous show abundant presence of Cp & Tf, both of which migrate faster upon deglycosylation (lanes 7 & 9). No reactivity for TfR or ferritin is detected in these samples (lanes 6–9). Human brain lysate was processed in parallel as a positive control (lane 5). Gapdh served as a loading control. (Cor: cornea; Ft: ferritin). (B) Relative distribution of iron modulating proteins within each tissue shows higher expression of TfR relative to Cp & ferritin in the cornea, & higher levels of Cp relative to the TfR & ferritin in the CB. (C) Quantitative comparison of protein expression by densitometry shows significantly higher levels of ferritin & Cp, & lower levels of TfR in the CB relative to the cornea. All values were normalized to Gapdh that provided the loading control. Values represent fold change ± SEM of the indicated n. (D) Probing of Western blots of bovine cornea & CB for Fpn revealed increased expression of Fpn (3.2 fold) in CB relative to the cornea (lanes 1–4). Lysates from human brain & bovine retina were analyzed in parallel as controls (lanes 5 & 6). Gapdh served as a loading control. (E) Quantification by densitometry shows 3.2 fold higher levels of Fpn in the CB relative to the cornea. Values are mean + SEM of the indicated n. **p < 0.01. The full images of the cropped blots have been provided in the Supplementary Data. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29859760), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-21502_rabbit-polyclonal-ferroportin-slc40a1-antibody-310202416171438.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - Effects of FANCD2 knockdown on levels of cell cycle regulators. Caki-1 cells were transfected with control siRNA (siCon) or FANCD2 siRNA (siFD2#2) and treated with AICAR for 24 h. The levels of FANCD2, p15Ink4B, and p21Cip1 were evaluated by immunoblotting. Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1002/2211-5463.12185) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/FANCD2-Antibody-Knockdown-Validated-NB100-182-img0040.jpg "<b>Biological Strategies Validation and Genetic Strategies Validation.</b>Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - Effects of FANCD2 knockdown on levels of cell cycle regulators. Caki-1 cells were transfected with control siRNA (siCon) or FANCD2 siRNA (siFD2#2) and treated with AICAR for 24 h. The levels of FANCD2, p15Ink4B, and p21Cip1 were evaluated by immunoblotting. Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1002/2211-5463.12185) licensed under a CC-BY license.")
![<b>Biological Strategies Validation and Genetic Strategies Validation.</b>Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - Resistance of FANCA-mutant cells with defective FANCD2 function to hydrogen peroxide (H2O2). Representative images illustrating staining for subnuclear FANCD2 foci in isogenic fibroblast pairs, either deficient for FANCD2 (PD20) or FANCA (PD220) and their respective wild-type (wt) complemented counterparts. Foci were visualized three hours after treatment with H2O2 (25 uM for 2 hours). Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/bmri/2008/821529/) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/FANCD2-Antibody-Knockdown-Validated-NB100-182-img0035.jpg)
![<b>Biological Strategies Validation. </b>Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - AMPK-activating AICAR treatment activates FANCD2, a pivotal molecule of Fanconi anemia DNA damage signaling pathway. AICAR treatment induces FANCD2 monoubiquitination in transformed normal fibroblasts (GM00637I). GM00637I cells were treated with 1 mm 2-deoxyglucose, 0.25 mM AICAR, or 1 mm phenformin for 24 h. Lysates were subjected to western blotting with FANCD2 Antibody, phospho-AMPKalpha1 (T172), and AMPKalpha and beta-actin. In FANCD2 blots, the position of monoubiquitinated FANCD2 (Ub-FANCD2) is indicated by an arrow. Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1002/2211-5463.12185) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/FANCD2-Antibody-Western-Blot-NB100-182-img0039.jpg)
![Simple Western: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - Simple Western lane view shows a specific band for FANCD2 in 0.1 mg/ml of HeLa lysate using FANCD2 Antibody. This experiment was performed under reducing conditions using the 12-230 kDa separation system.](http://images.novusbio.com/fullsize/FANCD2-Antibody-Simple-Western-NB100-182-img0014.jpg)
![Immunocytochemistry/Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - Analysis using the Biotin conjugate of FANCD2 Antibody. FANCD2 colocalizes in vivo with another protein in SiHa cells after cell exposure to IR. Proliferating SiHa cells were exposed to 10 Gy of IR and double color immunofluorescence staining was performed after 8 h. Images were captured in a Kodak digital image system on a Leica fluorescence microscope.](http://images.novusbio.com/fullsize/FANCD2-Antibody-Immunocytochemistry-Immunofluorescence-NB100-182-img0025.jpg)
![<b>Biological Strategies Validation. </b>Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - AICAR-induced FANCD2 monoubiquitination is dependent on AMPK. Inhibitor of AMPK blocks AICAR-induced FANCD2 monoubiquitination in GM00637I normal fibroblasts. Cells were pretreated with 5 um of Compound C (an AMPK inhibitor) 1 h before treatment with 0.25 mm AICAR for 24 h. Cell lysates were subjected to immunoblotting with FANCD2 Antibody to visualize monoubiquitinated FANCD2 (Ub-ANCD2). Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1002/2211-5463.12185) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/FANCD2-Antibody-Western-Blot-NB100-182-img0037.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - The FA-BRCA Pathway Is Activated on the Sex Chromosomes during Meiosis. Schematic of the FA-BRCA pathway. FA proteins analyzed in this study are shown in color. Western blot analysis with three independent anti-FANCD2 antibodies (G33, E33, and Novus NB100-182 antibody: NB). K561R, PD20 cells expressing a mutated form of FANCD2 incapable of monoubiquitination; WT, PD20 cells complemented with wild-type FANCD2; Vector, PD20 cells containing empty vector. Image collected and cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S2211124716313298), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/FANCD2-Antibody-Western-Blot-NB100-182-img0044.jpg)
![<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - HES1K109E mutation does not impair MMC-induced HES1 foci formation. HeLa cells transfected with HA-tagged HES1 or HES1K109E coding vectors were processed for immunofluorescence 16 h following treatment with MMC (120 ng/ml). Cells were double-stained with anti-HA and anti-FANCD2 antibodies (mouse anti-HA at 1:500; rabbit anti-FANCD2, Novus Biologicals NB100-182 at 1:2000) followed with secondary antibodies (Goat anti-mouse Alexafluor-555, ThermoScientific A-32727 and goat anti-rabbit Alexafluor-488, ThermoScientific, A-11008). Cell nuclei were labelled with TO-PRO-3 Iodide stain (1:400, ThermoScientific, T3605). Cells were visualized at 100x magnification. Image collected and cropped by CiteAb from the following publication (//bmcresnotes.biomedcentral.com/articles/10.1186/s13104-018-3243-7), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/FANCD2-Antibody-Immunocytochemistry-Immunofluorescence-NB100-182-img0045.jpg)
![Immunohistochemistry: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - The tissue sections were incubated with a primary antibody cocktail of FANCD2 Antibody (NB100-182) at a dilution of 1:1000 & a monoclonal anti-Ki67 mouse antibody at a dilution of 1:150 for 1 h at room temperature. Then the slides were incubated with a secondary antibody cocktail containing FITC conjugated anti-rabbit IgG and Alexafluor 594 donkey anti-mouse secondary. The sections were mounted on glass slides using a DAPI-containing embedding medium (Vysis Dapi 1, Abbott Laboratories, Downers Grove, IL, USA), then analyzed under a fluorescence microscope. FANCD2 (top) foci positive NSCL tumor, and FANCD2 (bottom)foci negative NSCL tumor. Magnification: 1000x. Image collected and cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fonc.2014.00368/abstract), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/FANCD2-Antibody-Immunohistochemistry-NB100-182-img0046.jpg)
![Western Blot: FANCD2 Antibody [NB100-182] - Analysis of FANCD2 (Molecular weight: 164.1 KDa) using the HRP conjugate of FANCD2 Antibody (lot C) in HeLa WCE.](http://images.novusbio.com/fullsize/FANCD2-Antibody-Western-Blot-NB100-182-img0029.jpg)
![Western Blot: FANCD2 Antibody [NB100-182] - Functional assessment of Fanconi anemia pathway. Western blot with FANCD2 Antibody of non-FA control lymphoblasts (LCL), proband LCL-A-2017, proband LCL-B-2017, FANCB mutant (null) LCL, and FANCD2 mutant (null) LCL. Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1002/mgg3.350), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/FANCD2-Antibody-Western-Blot-NB100-182-img0043.jpg)
![Immunocytochemistry/Immunofluorescence: FANCD2 Antibody [NB100-182] - Mouse Embryonic Fibroblasts Untreated and treated with Mitomycin C for 24hr with FANCD2 Antibody diluted 1:500 in 15%BCS+0.3% Triton X 100 in PBS. This image was submitted via customer Review.](http://images.novusbio.com/fullsize/FANCD2-Antibody-Immunocytochemistry-Immunofluorescence-NB100-182-img0031.jpg)
![Immunohistochemistry: FANCD2 Antibody [NB100-182] - Staining of human prostate, glandular epithelium using FANCD2 Antibody. Image using the Biotin format of this antibody.](http://images.novusbio.com/fullsize/FANCD2-Antibody-Immunohistochemistry-NB100-182-img0041.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - Western blot analysis of FANCD2 & ERCC1 expression in lung cancer cell H1299 following treatment with veliparib. PAR protein level was reduced at 6 h post veliparib 5 μM exposure & maintained at low levels through 48 h in the H1299 cells. However, FANCD2 & ERCC1 protein expression raised simultaneously in these cells post treatment with veliparib. The data suggest that veliparib induces compensatory activation of alternative DNA-repair pathways. C is control (without treatment). Image collected & cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fonc.2014.00368/abstract), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-310202415392537.jpg)
![Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - Workflow for MS-based quantification of CFS associated proteins. (A) Experimental workflow for SILAC-based quantitative MS identification of CFS associated proteins. (B) Flow cytometry analysis of cell cycle distribution of SILAC labeled cells used for enrichment of CFSs as illustrated in (A). (C) IF of FANCD2 & EdU incorporation to assess formation of FANCD2 foci at late replicating regions with & without APH treatments. Cells were synchronized as in (A) & (B). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31180492), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-310202415291919.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - The response to replication stress is a defining feature for Ewing sarcoma compared to the PHATE_1-high developmental context. (A) Western blot showing expression of Fanconi Anemia proteins FANCA, FANCD2, & FANCI in TC32 (Ewing sarcoma cell line) compared to IMR90 (fibroblast cell line); (B) Bar-plot showing the impact of FANC genes knockdown on cell viability in TC32 compared to IMR90; (C) Western blot showing the expression of FEN1 in Ewing sarcoma cell lines TC32, EWS502, & CHLA10 compared to IMR90; (D) Line plot showing the viability of Ewing sarcoma cells compared to IMR90 with increasing doses of FEN1 inhibitor. (** p ≤ 0.01; *** p ≤ 0.001; **** p ≤ 0.0001). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32290418), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-310202415334936.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - Activation of FANCD2 monoubiquitination following treatment with the EZH2 inhibitor EPZ-6438(A-C) MCF10A (A), HCT116 p53+/+ & p53-/- (B), & U2OS (C) cells were treated with the indicated concentrations of the EZH2-specific inhibitor EPZ-6438 for 24 h (A & B) or 24, 48, & 72 h (C). Whole-cell lysates were prepared & immunoblotted with the indicated antibodies. L.E., light exposure; D.E., dark exposure; L:S Ratio, ratio of monoubiquitinated to nonubiquitinated FANCI; or FANCD2 RBI, relative band intensity. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29100324), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-31020241539742.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - Inhibition of class I & II HDACs attenuates FANCD2 & FANCI monoubiquitination in BJ-TERT cells(A) & (B), HeLa cells were pre-treated with the indicated concentrations of trichostatin A (TSA) (A) or vorinostat (SAHA) (B) for 4 h, followed by co-incubation with (+) & without (-) 200 nM MMC for a further 20 h. Whole-cell lysates were prepared & immunoblotted with anti-FANCD2, anti-FANCI, anti-CHK1 pS345, anti-CHK2 pT68, anti-H4K16ac, & anti-alpha -Tubulin antibodies. (C) & (D), BJ-TERT cells were treated identically to that described for HeLa cells above. L:S Ratio, ratio of monoubiquitinated to nonubiquitinated FANCD2 or FANCI. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29100324), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-31020241535664.jpg)
![Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - Representative images of the cellular response to RS (induced by a low dose of hydroxyurea; HU) in U2OS cells, as determined by IF staining for gamma H2AX & FANCD2A selected cell is defined by the yellow arrow, & is enlarged in the bottom panel. Scale bars are indicated. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.16940), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-310202415395917.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - Functional evaluation of FANCBWT & mutant cDNA. Proband fibroblasts, FANCB mutant (null) fibroblasts & BJ control fibroblasts were HPV16 E6E7 transformed. Either empty vector, wild‐type FANCBcDNA, or mutant FANCBcDNA (p.A319*) was introduced into proband fibroblasts & FANCB mutant (null) fibroblasts. After puromycin selection, cells were cultured with or without MMC 1 μM for 24 hr, after which cells were harvested for the FANCD2 & HA western blot assays Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29193904), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-310202415392569.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - BRD4770-induced activation of the FA pathway may occur via inhibition of the PRC2 complex(A) U2OS cells were incubated in the absence (NT) or presence of BRD4770, DZNep, UNC0646, & DZNep & UNC0646 combined (4 μM each) for 24 h. Whole-cell lysates were prepared & immunoblotted with anti-FANCD2, anti-FANCI, anti-EZH2, & anti-alpha -Tubulin antibodies. (B) U2OS cells were incubated in the absence or presence of 2, 5, & 10 μM BRD4770 for 24, 48, or 72 h. Whole-cell lysates were prepared & immunoblotted with anti-FANCD2, anti-FANCI, anti-CHK1 pS345, anti-EZH2, anti-H3K27me3, & anti-alpha -Tubulin antibodies. L:S Ratio, ratio of monoubiquitinated to nonubiquitinated FANCI; or FANCD2 RBI, relative band intensity. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29100324), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-3102024153434.jpg)
![Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - Chemical library screening for small molecules that inhibit the Fanconi anemia pathway. (A) Schematic of the screening for small molecules that inhibit IR-induced FANCD2 foci formation. (B) Representative photomicrographs of EGFP-FANCD2 foci in PD20F-EGFP-FANCD2 cells untreated & treated with the indicated compounds at the indicated concentration. The cells were treated with compounds immediately before irradiation (15 Gy), & fixed after 12 hours. (C) FANCD2 foci in HeLa cells untreated & treated with the indicated compounds at the indicated concentration. The cells were fixed 8 hours after irradiation (10 Gy) & immunostained with anti-FANCD2 antibody. Scale bar = 20 μm. Image collected & cropped by CiteAb from the following publication (//molecular-cancer.biomedcentral.com/articles/10.1186/1476-4598-11-26), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-310202415345333.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - AMPK‐activating AICAR treatment activates FANCD2, a pivotal molecule of Fanconi anemia DNA damage signaling pathway. (A) AICAR treatment induces FANCD2 monoubiquitination in transformed normal fibroblasts (GM00637I). GM00637I cells were treated with 1 mm 2‐deoxyglucose, 0.25 mM AICAR, or 1 mm phenformin for 24 h. Lysates were subjected to western blotting with anti‐FANCD2, phospho‐AMPK alpha 1 (T172), & AMPK alpha & beta ‐actin. In FANCD2 blots, the position of monoubiquitinated FANCD2 (Ub‐FANCD2) is indicated by an arrow. (B) AICAR treatment induces formation of FANCD2 nuclear foci in GM00637I fibroblasts. Cells grown on coverslips in 12‐well plates were treated with 0.25 mm AICAR for 24 h. Cells were immunostained with FANCD2 antibody & Alexa 488‐conjugated anti‐rabbit secondary antibody. FANCD2 foci were visualized by confocal microscopy. Representative images are shown at the top. The number of foci per cell was counted & plotted for ≥ 20 cells (bottom panel). The values represent the mean ± SEM (Student's t‐test, ***P 5 gamma H2AX foci were scored. At least 300 nuclei were scored for each treatment & this experiment was performed three times with similar results. Error bars represent the standard errors of the means from three independent experiments. (B) BJ-TERT cells were incubated with (+) & without (-) 0.4 μM etoposide (VP-16), in the absence or presence of 10 μM BRD4770, 5 μM GSK-J1 & 2.5 μM BIX01294, for 24 h. Whole-cell lysates were immunoblotted with anti-gamma H2AX & anti-PCNA (loading control) antibodies. (C) U2OS cells were incubated in the absence (NT) or presence of 200 nM MMC or 5 & 10 μM BRD4770 for 24 h. Whole-cell lysates were prepared & immunoblotted with anti-FANCD2, anti-FANCA, anti-UBE2T, anti-USP1, anti-CHK1 pS345, anti-RPA, anti-RPA pS4/8, & anti-alpha -Tubulin antibodies. L:S Ratio, ratio of monoubiquitinated to nonubiquitinated FANCD2; RBI, relative band intensity. (D) U2OS cells were incubated in the absence or presence of 2, 5, & 10 μM BRD4770 for 24, 48, or 72 h. Cells were fixed in ice-cold ethanol, stained with propidium iodide, & analyzed by flow cytometry. Cell cycle stage distributions were determined using FlowJo v10.2. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29100324), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-310202415525240.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - AICAR‐induced FANCD2 monoubiquitination is dependent on AMPK. (A) Inhibitor of AMPK blocks AICAR‐induced FANCD2 monoubiquitination in GM00637I normal fibroblasts. Cells were pretreated with 5 μm of Compound C (an AMPK inhibitor) 1 h before treatment with 0.25 mm AICAR for 24 h. Cell lysates were subjected to immunoblotting with FANCD2 to visualize monoubiquitinated FANCD2 (Ub‐FANCD2). (B) AMPK inhibitor abrogates AICAR‐induced FANCD2 nuclear foci formation in GM00637I fibroblasts. GM006387I cells grown on coverslips were pretreated with 5 μm Compound C 1 h prior to 0.25 mm AICAR treatment for 24 h. FANCD2 foci were visualized by immunofluorescence staining & confocal microscopy. Representative images are shown at the top. The number of foci per cell was counted & plotted for ≥ 20 cells (bottom panel). The values represent the mean ± SEM (Student's t‐test, *P < 0.05; ***P < 0.001). (C) Knockdown of AMPK alpha 1 inhibits AICAR‐induced FANCD2 monoubiquitination in Caki‐1 cells. Caki‐1 cells were transfected with siRNAs (siControl or siAMPK#8) & after 48 h, AICAR was treated for 24 h. FANCD2 monoubiquitination was monitored by immunoblotting. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28174693), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-310202415533833.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - LNK depletion restores FA-like human progenitor cells. a depicts a schematic overview of isolation of primary human HSPCs for lentiviral transduction followed by CFC assay. b UCB-derived CD34 + cells were sequentially infected with lentiviruses expressing shRNA to Luciferase (Luc) or FANCD2 (D2) with GFP marker, followed by shRNA to Luc or LNK with mCherry marker. GFP + mCherry + cells were then sorted & plated onto semi-solid methylcellulose media. Colony forming progenitor numbers are shown with a two-tailed Student's t-test. Representative data from two independent repeats are shown. c TF-1 cells with shRNA-mediated depletion of LNK or FANCD2 or CRISPR/Cas9-mediated depletion of LNK were analyzed for depletion efficiency by western blot. Luc, luciferase. gRNA, guide RNA Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30254368), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-310202415532063.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - SAN1 functions independently of the FA pathway & does not affect FA pathway activation. a, b CSAs of HeLa WT & SAN1−/− cells treated with scrambled ctrl siRNA or FANCD2 siRNA, in response to Cisplatin & MMC (N = 3). Statistical significance determined by two-way ANOVA. Error bars denote s.e.m. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. c Immunoblot showing siRNA knockdown of FANCD2 in HeLa WT & SAN1−/− cells. d IF staining of FANCD2 foci in HeLa WT cells & SAN1−/− cells treated with 0.045 μM MMC. e Immunoblot of FANCD2 showing mono-ubiquitylation in HeLa WT & SAN1−/− cells treated with vehicle or 0.045 μM MMC. f, g CSAs of HeLa WT & SAN1−/− cells treated with ctrl or SNM1A siRNA & exposed to Cisplatin or MMC. Statistical significance was determined by two-way ANOVA test. h Immunoblot of SNM1A in HeLa WT & SAN1−/− cells treated with ctrl or SNM1A siRNA Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29968717), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-31020241553749.jpg)
![Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - SAN1 functions independently of the FA pathway & does not affect FA pathway activation. a, b CSAs of HeLa WT & SAN1−/− cells treated with scrambled ctrl siRNA or FANCD2 siRNA, in response to Cisplatin & MMC (N = 3). Statistical significance determined by two-way ANOVA. Error bars denote s.e.m. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. c Immunoblot showing siRNA knockdown of FANCD2 in HeLa WT & SAN1−/− cells. d IF staining of FANCD2 foci in HeLa WT cells & SAN1−/− cells treated with 0.045 μM MMC. e Immunoblot of FANCD2 showing mono-ubiquitylation in HeLa WT & SAN1−/− cells treated with vehicle or 0.045 μM MMC. f, g CSAs of HeLa WT & SAN1−/− cells treated with ctrl or SNM1A siRNA & exposed to Cisplatin or MMC. Statistical significance was determined by two-way ANOVA test. h Immunoblot of SNM1A in HeLa WT & SAN1−/− cells treated with ctrl or SNM1A siRNA Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29968717), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-3102024165713.jpg)
![Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - The FANCD2 NLS is required for the nuclear localization of a subset of FANCI.(A) FA-D2 patient cells or FA-D2 cells stably expressing FANCD2-WT were incubated in the absence (NT) or presence of MMC for 24 h, fixed, stained with rabbit polyclonal anti-FANCD2 or anti-FANCI antibody & counterstained with phalloidin & DAPI. AF-488, Alexa Fluor 488. (B) FA-D2 cells stably expressing LacZ, FANCD2-WT, FANCD2-∆N57, FANCD2-∆N100, & FANCD2-3N were incubated in the absence (NT) or presence of MMC for 24 h, fixed, & stained with rabbit polyclonal anti-FANCI antibody, & counterstained with phalloidin & DAPI. At least 300 cells were scored for cytoplasmic (Cyto.), nuclear (Nucl.), & both cytoplasmic & nuclear (Both) localization of FANCI. (C) COS-7 cells were transiently transfected with no DNA, FANCI-GFP, FANCI-GFP plus FANCD2-V5-WT, or FANCI-GFP plus FANCD2-V5-∆N57. Whole-cell lysates were immunoprecipitated with anti-V5 or anti-GFP antibodies & immune complexes immunoblotted with anti-GFP & anti-V5 antibodies. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24278431), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-3102024161268.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - Functional assessment of Fanconi anemia pathway. (a) The experimental scheme for MMC treatment. Twenty‐four hours after plating, cells were cultured with or without MMC 1 μM for an additional 24 hr, after which the cells were harvested for western blot or immunostaining. (b) Western blot with FANCD2 antibody of BJ, proband fibroblasts, FANCB mutant (null) fibroblasts & FANCD2 mutant (null) fibroblasts. (c) Western blot with FANCD2 antibody of non‐FA control lymphoblasts (LCL), proband LCL‐A‐2017, proband LCL‐B‐2017, FANCB mutant (null) LCL, & FANCD2 mutant (null) LCL. (d) Representative figures of FANCD2 foci formation in the indicated cells. (e) Quantification of FANCD2 foci formation following treatment with or without 1 μM MMC. Experiments were performed in triplicate. One hundred cells were counted for each experiment Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29193904), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-310202415525261.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - The FAN1 PIP-box motif is not required for FAN1 foci formation upon exposure to MMC. a Total cell extracts & chromatin-enriched fractions of U2OS cells expressing the indicated eGFP-FAN1 variants, treated or mock-treated with MMC (150 ng/ml, 24 h), were analysed by immunoblotting using the indicated antibodies. A representative blot of three independent experiments is shown. b Cells as in a were immunostained with anti-FANCD2 antibody. Representative images are shown. Scale bar: 25 μm. c, d Quantification of eGFP-FAN1 foci count (c) & the sum of their intensities (d) was obtained from QIBC analysis of b. Median levels are indicated by black bars. Statistical analyses were carried out using unpaired, two-tailed t-tests. P values expressed as ***(P < 0.01) were considered significant, n = 3. e Total cell extracts derived from cells as in a, treated or mock-treated with MMC (150 ng/ml, 24 h), were incubated with anti-eGFP affinity resin. Inputs & immunoprecipitates were analysed by immunoblotting with the indicated antibodies. A representative blot of two independent experiments is shown Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29051491), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-31020241682111.jpg)
![Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - Functional assessment of Fanconi anemia pathway. (a) The experimental scheme for MMC treatment. Twenty‐four hours after plating, cells were cultured with or without MMC 1 μM for an additional 24 hr, after which the cells were harvested for western blot or immunostaining. (b) Western blot with FANCD2 antibody of BJ, proband fibroblasts, FANCB mutant (null) fibroblasts & FANCD2 mutant (null) fibroblasts. (c) Western blot with FANCD2 antibody of non‐FA control lymphoblasts (LCL), proband LCL‐A‐2017, proband LCL‐B‐2017, FANCB mutant (null) LCL, & FANCD2 mutant (null) LCL. (d) Representative figures of FANCD2 foci formation in the indicated cells. (e) Quantification of FANCD2 foci formation following treatment with or without 1 μM MMC. Experiments were performed in triplicate. One hundred cells were counted for each experiment Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29193904), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-3102024155378.jpg)
![Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - DNA damage & cell survivalas a function of FANCD2 status. (a) Representative images of the formation of gamma H2AX foci in PD20 versus wild-typecomplemented cells 30 minutes after completion of H2O2 treatment. (b) Quantification of gamma H2AX foci response. Data represent means with upper standard error based on two independent experiments. (c) G2-type chromosomal aberrations are expressed as breaks per cell as a function of increasing H2O2 concentration in FANCD2-deficient andwild-type complemented PD20 cells. Data represent means with SEM based on at least three repeat experiments. (d) Apoptosis induction by H2O2 (50 μM) in cells with or without wild-type FANCD2 using fluorescence microscopy to assess apoptotic morphology by DAPI staining & flow cytometric analysis for sub-G1 DNA content. Representative experiments based on theapoptotic response at 24 hours are shown (similar results were obtained at 48 hours & with 25 μM H2O2). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/18483568), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-31020241682179.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - The FANCD2 NLS is required for efficient FANCD2 & FANCI monoubiquitination & chromatin association.(A) FA-D2 cells stably expressing LacZ, FANCD2-WT, FANCD2-K561R, FANCD2-∆N57, FANCD2-∆N100 & FANCD2-3N were incubated in the absence & presence of 250 nM MMC for 18 h, & whole-cell lysates were immunoblotted with antibodies to FANCD2, V5, FANCI & alpha -tubulin. The FANCD2 & FANCI L/S ratios are the ratios of monoubiquitinated to nonubiquitinated protein, & were calculated by measuring protein band intensities using ImageJ image processing & analysis software (//rsb.info.nih.gov/ij/). (B & C) FA-D2 cells stably expressing FANCD2-WT, FANCD2-∆N57, FANCD2-∆N100 & FANCD2-3N were treated as above & cell pellets were fractionated into soluble (S) & chromatin-associated (C) fractions. Fractions were immunoblotted with antibodies against V5, FANCI, alpha -tubulin & H2A. W, unfractionated whole cell extract. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24278431), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-310202415535141.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - AICAR‐induced FANCD2 monoubiquitination requires FANCA. (A) AICAR‐induced FANCD2 monoubiquitination does not occur in transformed FANCA−/− fibroblasts originated from a patient with defective FANCA. Transformed FANCA−/− (GM06914B) or normal fibroblasts (GM00637I) were treated with 0.25 mm AICAR for 24 h, & monoubiquitinated FANCD2 (Ub‐FANCD2) was visualized by immunoblotting. (B) AICAR‐induced FANCD2 nuclear foci formation is abrogated in FANCA−/− fibroblasts. FANCD2 nuclear foci were visualized by immunofluorescence staining & confocal microscopy. (C) Knockdown of FANCA abolishes FANCD2 monoubiquitination after AICAR treatment in Caki‐1 cells. Caki‐1 cells were transfected with FANCA‐targeting siRNAs (siFANCA#5 or siFANCA#6), treated with AICAR & subjected to FANCD2 immunoblotting. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28174693), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-31020241612650.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - Acetaldehyde toxicity to human FANCD2‐deleted human cells & FANCD2 ubiquitylation in BRCA2‐deleted cellsA, BHuman DLD1 cells in which FANCD2 was deleted with CRISPR/Cas9 & control cells were incubated with the indicated concentrations of cisplatin (A) or acetaldehyde (B) for 6 days before processing for dose‐dependent viability assays. Graphs are representative of two independent experiments, each performed in triplicate. Error bars represent SD of triplicate values obtained from a single experiment. Inset, Western blot detection of FANCD2 expression. SMC1 was used as a loading control.CBRCA2‐proficient (+BRCA2) or BRCA2‐deficient (−BRCA2) DLD1 cells were incubated with 4 mM acetaldehyde for 48 h before being processed for immunoblotting as indicated.DH1299 cells expressing a DOX‐inducible BRCA2 shRNA were grown in the presence or absence of DOX & transfected with control or FANCD2 siRNA before being processed for immunoblotting as indicated. DOX, doxycycline. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28729482), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-310202415533832.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - Functional assessment of Fanconi anemia pathway. (a) The experimental scheme for MMC treatment. Twenty‐four hours after plating, cells were cultured with or without MMC 1 μM for an additional 24 hr, after which the cells were harvested for western blot or immunostaining. (b) Western blot with FANCD2 antibody of BJ, proband fibroblasts, FANCB mutant (null) fibroblasts & FANCD2 mutant (null) fibroblasts. (c) Western blot with FANCD2 antibody of non‐FA control lymphoblasts (LCL), proband LCL‐A‐2017, proband LCL‐B‐2017, FANCB mutant (null) LCL, & FANCD2 mutant (null) LCL. (d) Representative figures of FANCD2 foci formation in the indicated cells. (e) Quantification of FANCD2 foci formation following treatment with or without 1 μM MMC. Experiments were performed in triplicate. One hundred cells were counted for each experiment Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29193904), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-310202415525258.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - SAN1 functions independently of the FA pathway & does not affect FA pathway activation. a, b CSAs of HeLa WT & SAN1−/− cells treated with scrambled ctrl siRNA or FANCD2 siRNA, in response to Cisplatin & MMC (N = 3). Statistical significance determined by two-way ANOVA. Error bars denote s.e.m. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. c Immunoblot showing siRNA knockdown of FANCD2 in HeLa WT & SAN1−/− cells. d IF staining of FANCD2 foci in HeLa WT cells & SAN1−/− cells treated with 0.045 μM MMC. e Immunoblot of FANCD2 showing mono-ubiquitylation in HeLa WT & SAN1−/− cells treated with vehicle or 0.045 μM MMC. f, g CSAs of HeLa WT & SAN1−/− cells treated with ctrl or SNM1A siRNA & exposed to Cisplatin or MMC. Statistical significance was determined by two-way ANOVA test. h Immunoblot of SNM1A in HeLa WT & SAN1−/− cells treated with ctrl or SNM1A siRNA Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29968717), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-3102024161265.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - AICAR‐induced FANCD2 monoubiquitination requires FANCA. (A) AICAR‐induced FANCD2 monoubiquitination does not occur in transformed FANCA−/− fibroblasts originated from a patient with defective FANCA. Transformed FANCA−/− (GM06914B) or normal fibroblasts (GM00637I) were treated with 0.25 mm AICAR for 24 h, & monoubiquitinated FANCD2 (Ub‐FANCD2) was visualized by immunoblotting. (B) AICAR‐induced FANCD2 nuclear foci formation is abrogated in FANCA−/− fibroblasts. FANCD2 nuclear foci were visualized by immunofluorescence staining & confocal microscopy. (C) Knockdown of FANCA abolishes FANCD2 monoubiquitination after AICAR treatment in Caki‐1 cells. Caki‐1 cells were transfected with FANCA‐targeting siRNAs (siFANCA#5 or siFANCA#6), treated with AICAR & subjected to FANCD2 immunoblotting. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28174693), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-31020241553513.jpg)
![Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - BRCA2-deficient cells accumulate single-stranded DNA lesions in G2. a gamma H2AX foci analysis in early mitotic cells. Cells were untreated, or treated with RO-3306 (10 µM, 24 h) to delay mitotic entry, & released for 1 h before analysis of gamma H2AX & FANCD2 foci pairs in early mitotic cells. Representative images are shown (left). Analysis is by an unpaired two-tailed t-test. n = 3. Scale bars 10 µm. b–g Cells treated with the indicated siRNAs or mirin (50 µM, 5 h) were incubated with EdU for 30 min & then analyzed for gamma H2AX & RPA foci in G2 cells (EdU–, 2N DNA content). Representative deconvolved images are shown (b). Quantification of gamma H2AX foci (top, c–g) & RPA+ gamma H2AX foci (bottom, c–g) for BRCA2-deficient cells (c), cells transfected with siRNAs (d, RAD51; e SMARCAL1; f EXO1 & DNA2), & cells treated with mirin (g) are shown. n ≥ 3. Scale bars 10 µm. h, i Cells were incubated with EdU as in b before analysis of gamma H2AX & pCHK2-T68 (h) or pATM-S1981 (i) foci in G2 cells. Representative deconvolved images with magnified inset highlighting foci co-localization are shown (left in each panel). n ≥ 3. Scale bars 10 µm. Error bars s.d. ns, not significant; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001 (unpaired two-tailed t-test) Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-017-00634-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-31020241612682.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - SAN1 functions independently of the FA pathway & does not affect FA pathway activation. a, b CSAs of HeLa WT & SAN1−/− cells treated with scrambled ctrl siRNA or FANCD2 siRNA, in response to Cisplatin & MMC (N = 3). Statistical significance determined by two-way ANOVA. Error bars denote s.e.m. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. c Immunoblot showing siRNA knockdown of FANCD2 in HeLa WT & SAN1−/− cells. d IF staining of FANCD2 foci in HeLa WT cells & SAN1−/− cells treated with 0.045 μM MMC. e Immunoblot of FANCD2 showing mono-ubiquitylation in HeLa WT & SAN1−/− cells treated with vehicle or 0.045 μM MMC. f, g CSAs of HeLa WT & SAN1−/− cells treated with ctrl or SNM1A siRNA & exposed to Cisplatin or MMC. Statistical significance was determined by two-way ANOVA test. h Immunoblot of SNM1A in HeLa WT & SAN1−/− cells treated with ctrl or SNM1A siRNA Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29968717), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-31020241683925.jpg)
![Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - BRCA2 deficiency causes DNA under replication that results in abnormal mitoses. a–cBRCA2∆Ex3-4/– cells released for 22 h from serum starvation to increase mitotic cells, incubated w/ EdU for 1 h, & then analyzed for mitotic DNA synthesis. Early mitotic cells defined as being in prophase, prometaphase,/metaphase analyzed for EdU foci that co-localize w/ FANCD2 foci pairs. a Representative images of mitotic DNA synthesis. Scale bars 10 µm. b Percent early mitotic cells containing EdU foci, analyzed by an unpaired two-tailed t-test. n = 3. c FANCD2 foci pairs w//w/out EdU foci co-localization. Graphs represent pooled results of 3 independent experiments, each analyzed by a two-tailed Mann–Whitney test. Median FANCD2 foci pair number, red bars. Image collected & cropped by CiteAb from following publication (//www.nature.com/articles/s41467-017-00634-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-310202415541981.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - The FANCD2 NLS is required for efficient FANCD2 & FANCI monoubiquitination & chromatin association.(A) FA-D2 cells stably expressing LacZ, FANCD2-WT, FANCD2-K561R, FANCD2-∆N57, FANCD2-∆N100 & FANCD2-3N were incubated in the absence & presence of 250 nM MMC for 18 h, & whole-cell lysates were immunoblotted with antibodies to FANCD2, V5, FANCI & alpha -tubulin. The FANCD2 & FANCI L/S ratios are the ratios of monoubiquitinated to nonubiquitinated protein, & were calculated by measuring protein band intensities using ImageJ image processing & analysis software (//rsb.info.nih.gov/ij/). (B & C) FA-D2 cells stably expressing FANCD2-WT, FANCD2-∆N57, FANCD2-∆N100 & FANCD2-3N were treated as above & cell pellets were fractionated into soluble (S) & chromatin-associated (C) fractions. Fractions were immunoblotted with antibodies against V5, FANCI, alpha -tubulin & H2A. W, unfractionated whole cell extract. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24278431), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-31020241612699.jpg)
![Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - Detection of FANCD2 foci formation in human lung tumors by the FATSI staining analysis. The paraffin-embedded lung tumor tissues sections were deparaffinized & rehydrated. The tissue sections were incubated with a primary antibody cocktail of rabbit polyclonal FANCD2 antibody (Novus Biologicals, Littleton, CO, USA) at a dilution of 1:1000 & a monoclonal anti-Ki67 mouse antibody (Dako, Carpenteria, CA, USA) at a dilution of 1:150 for 1 h at room temperature. Sections then were incubated with a secondary antibody cocktail containing FITC conjugated anti-rabbit IgG & Alexafluor 594 donkey anti-mouse secondary for 1 h at room temperature. The sections were mounted on glass slides using a 4′ 6-diamidino-2-phenylindole (DAPI)-containing embedding medium (Vysis Dapi 1, Abbott Laboratories, Downers Grove, IL, USA). The slides were analyzed under a fluorescence microscope. (A) FANCD2 foci positive NSCL tumor, & (B) FANCD2 foci negative NSCL tumor. Magnification: 1000×. Image collected & cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fonc.2014.00368/abstract), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-310202415541977.jpg)
![Immunocytochemistry/ Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - Normal DNA damage responses in FANCD2-deficient cells treated with hydrogen peroxide (H2O2). (a) Illustration of RAD51 foci formation in PD20 & PD20-wtD2 cells 5 hours after exposure to MMC (0.25 μg/mL for 1 hour) or H2O2 (25 μM for 2 hours). (b) Induction of RAD51 foci formation above background levels by MMC or H2O2. Because equal drug concentrations were used, rather than isoeffective concentrations with regard to cell survival, the extent of foci induction between FANCD-deficient & -complemented cells is not directly comparable. Data represent means with upperstandard error based on three independent repeats. (c) DNA synthesis measured by BrdU pulse labeling in cells treated with ionizing radiation (IR, 8 Gy) or H2O2 (25 μM). Data represent means with upper standard error based on twoindependent experiments. (d) Cell cycle distribution of propidium-iodide cell populations by flow cytometry. A representative experiment is shown. Percentages of cells in the G1, S, & G2 (and M) phases of the cell cycle are indicated. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/18483568), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-310202415541983.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - Dicer inhibition prevents FANCD2 foci formation after replication stress without affecting FANCD2 mono-ubiquitylation.(A) HCT116 cells were treated with control or Dicer siRNA & then with aphidicolin to induce replicative stress. They were then stained for FANCD2 & analyzed by immunofluorescence microscopy. (B) Quantification of the experiments in panel (A) showing the percentage of FANCD2 positive cells. Error bars represent the SD of three independent experiments. To score the FANCD2 positive cells a threshold value was calculated on the base of the average number of foci in control cells for each replicate, using Image J. Unpaired t-test: *p < 0.05. (C) FANCD2 foci formation in WT & Dicer Ex5/Ex5 cells after replication stress induced by aphidicolin. (D) Quantification of the experiments in panel (C) showing the percentage of FANCD2 positive cells. Error bars represent the SD of three independent experiments. Unpaired t-test: **p < 0.005. (E) Western blotting showing the levels of FANCD2 mono-ubiquitylation (L-FANCD2) after siRNA-mediated inhibition of Dicer, in the presence or absence of replication stress induced by aphidicolin. Vinculin was used as a loading control. (F) Western blotting showing FANCD2 mono-ubiquitylation levels after Drosha inhibition by siRNA. (G) FANCD2 mono-ubiquitylation levels in WT & Dicer Ex5/Ex5 cells assayed by western blotting. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31320994), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-310202415532033.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - Acetaldehyde toxicity to human FANCD2‐deleted human cells & FANCD2 ubiquitylation in BRCA2‐deleted cellsA, BHuman DLD1 cells in which FANCD2 was deleted with CRISPR/Cas9 & control cells were incubated with the indicated concentrations of cisplatin (A) or acetaldehyde (B) for 6 days before processing for dose‐dependent viability assays. Graphs are representative of two independent experiments, each performed in triplicate. Error bars represent SD of triplicate values obtained from a single experiment. Inset, Western blot detection of FANCD2 expression. SMC1 was used as a loading control.CBRCA2‐proficient (+BRCA2) or BRCA2‐deficient (−BRCA2) DLD1 cells were incubated with 4 mM acetaldehyde for 48 h before being processed for immunoblotting as indicated.DH1299 cells expressing a DOX‐inducible BRCA2 shRNA were grown in the presence or absence of DOX & transfected with control or FANCD2 siRNA before being processed for immunoblotting as indicated. DOX, doxycycline. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28729482), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-31020241553733.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - The FAN1 PIP-box motif is not required for FAN1 foci formation upon exposure to MMC. a Total cell extracts & chromatin-enriched fractions of U2OS cells expressing the indicated eGFP-FAN1 variants, treated or mock-treated with MMC (150 ng/ml, 24 h), were analysed by immunoblotting using the indicated antibodies. A representative blot of three independent experiments is shown. b Cells as in a were immunostained with anti-FANCD2 antibody. Representative images are shown. Scale bar: 25 μm. c, d Quantification of eGFP-FAN1 foci count (c) & the sum of their intensities (d) was obtained from QIBC analysis of b. Median levels are indicated by black bars. Statistical analyses were carried out using unpaired, two-tailed t-tests. P values expressed as ***(P < 0.01) were considered significant, n = 3. e Total cell extracts derived from cells as in a, treated or mock-treated with MMC (150 ng/ml, 24 h), were incubated with anti-eGFP affinity resin. Inputs & immunoprecipitates were analysed by immunoblotting with the indicated antibodies. A representative blot of two independent experiments is shown Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29051491), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-3102024168393.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - AMPK‐activating AICAR treatment activates FANCD2, a pivotal molecule of Fanconi anemia DNA damage signaling pathway. (A) AICAR treatment induces FANCD2 monoubiquitination in transformed normal fibroblasts (GM00637I). GM00637I cells were treated with 1 mm 2‐deoxyglucose, 0.25 mM AICAR, or 1 mm phenformin for 24 h. Lysates were subjected to western blotting with anti‐FANCD2, phospho‐AMPK alpha 1 (T172), & AMPK alpha & beta ‐actin. In FANCD2 blots, the position of monoubiquitinated FANCD2 (Ub‐FANCD2) is indicated by an arrow. (B) AICAR treatment induces formation of FANCD2 nuclear foci in GM00637I fibroblasts. Cells grown on coverslips in 12‐well plates were treated with 0.25 mm AICAR for 24 h. Cells were immunostained with FANCD2 antibody & Alexa 488‐conjugated anti‐rabbit secondary antibody. FANCD2 foci were visualized by confocal microscopy. Representative images are shown at the top. The number of foci per cell was counted & plotted for ≥ 20 cells (bottom panel). The values represent the mean ± SEM (Student's t‐test, ***P < 0.001). (C) AICAR treatment induces FANCD2 monoubiquitination in Caki‐1 cells. Caki‐1 cells were treated with 0.25 mm AICAR for 24 h & monoubiquitination of FANCD2 was monitored as in (A). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28174693), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-310202415525212.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - AICAR‐induced FANCD2 monoubiquitination is dependent on AMPK. (A) Inhibitor of AMPK blocks AICAR‐induced FANCD2 monoubiquitination in GM00637I normal fibroblasts. Cells were pretreated with 5 μm of Compound C (an AMPK inhibitor) 1 h before treatment with 0.25 mm AICAR for 24 h. Cell lysates were subjected to immunoblotting with FANCD2 to visualize monoubiquitinated FANCD2 (Ub‐FANCD2). (B) AMPK inhibitor abrogates AICAR‐induced FANCD2 nuclear foci formation in GM00637I fibroblasts. GM006387I cells grown on coverslips were pretreated with 5 μm Compound C 1 h prior to 0.25 mm AICAR treatment for 24 h. FANCD2 foci were visualized by immunofluorescence staining & confocal microscopy. Representative images are shown at the top. The number of foci per cell was counted & plotted for ≥ 20 cells (bottom panel). The values represent the mean ± SEM (Student's t‐test, *P < 0.05; ***P < 0.001). (C) Knockdown of AMPK alpha 1 inhibits AICAR‐induced FANCD2 monoubiquitination in Caki‐1 cells. Caki‐1 cells were transfected with siRNAs (siControl or siAMPK#8) & after 48 h, AICAR was treated for 24 h. FANCD2 monoubiquitination was monitored by immunoblotting. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28174693), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-31020241553516.jpg)
![Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - Acetaldehyde toxicity to human FANCD2‐deleted human cells & FANCD2 ubiquitylation in BRCA2‐deleted cellsA, BHuman DLD1 cells in which FANCD2 was deleted with CRISPR/Cas9 & control cells were incubated with the indicated concentrations of cisplatin (A) or acetaldehyde (B) for 6 days before processing for dose‐dependent viability assays. Graphs are representative of two independent experiments, each performed in triplicate. Error bars represent SD of triplicate values obtained from a single experiment. Inset, Western blot detection of FANCD2 expression. SMC1 was used as a loading control.CBRCA2‐proficient (+BRCA2) or BRCA2‐deficient (−BRCA2) DLD1 cells were incubated with 4 mM acetaldehyde for 48 h before being processed for immunoblotting as indicated.DH1299 cells expressing a DOX‐inducible BRCA2 shRNA were grown in the presence or absence of DOX & transfected with control or FANCD2 siRNA before being processed for immunoblotting as indicated. DOX, doxycycline. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28729482), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-182_rabbit-polyclonal-fancd2-antibody-310202415541966.jpg)
![Western Blot: SLC6A3/DAT1 Antibody (mAb16) [NBP2-22164] - Western blot analysis of DAT protein. Band shown at approx. 75 kDa. Image submitted by verified customer review.](http://images.novusbio.com/fullsize/SLC6A3-DAT1-Antibody-mAb16-Western-Blot-NBP2-22164-img0005.jpg "Western Blot: SLC6A3/DAT1 Antibody (mAb16) [NBP2-22164] - Western blot analysis of DAT protein. Band shown at approx. 75 kDa. Image submitted by verified customer review.")
![Immunocytochemistry/Immunofluorescence: DAT1 Antibody (mAb16) [NBP2-22164] - The DAT1 antibody was tested in PC12 cells at a 1:250 dilution against DyLight 488 (Green). Actin and nuclei were counterstained with Phalloidin-AlexaFluor 568 (Red) and DAPI (Blue), respectively.](http://images.novusbio.com/fullsize/DAT1-Antibody-mAb16-Immunocytochemistry-Immunofluorescence-NBP2-22164-img0004.jpg)
![Immunohistochemistry-Paraffin: DAT1 Antibody (mAb16) [NBP2-22164] - IHC analysis of DAT1 in mouse brain using DAB with hematoxylin counterstain.](http://images.novusbio.com/fullsize/DAT1-Antibody-mAb16-Immunohistochemistry-Paraffin-NBP2-22164-img0002.jpg)
![Western Blot: DAT1 Antibody (mAb16) [NBP2-22164] - Analysis of DAT1 expression in rat striatal membrane tissue lysate.](http://images.novusbio.com/fullsize/DAT1-Antibody-mAb16-Western-Blot-NBP2-22164-img0003.jpg)