Enzyme-linked immunosorbent assay (ELISA) is an analytic biochemical assay that utilizes enzyme-linked antibodies on a microwell platform to quantitate specific proteins within biological samples. Visual detection is then achieved by adding the enzyme’s substrate, which causes a color-changing reaction to occur. There are three different types of ELISA that are most commonly used: indirect ELISA, sandwich ELISA and competitive ELISA. These assays are extremely useful for high-throughput diagnostic screenings drug testing.
Novus offers a range of support options for ELISA when you buy a product to help you with your ELISA experiment including:
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NB100-664SS | |||
| Azide and BSA Free | NBP2-81007 | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NBP1-47423SS | |||
| Azide and BSA Free | NBP2-80595 | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NB100-1614-0.02ml | |||
![N/A CXCL10/IP-10/CRG-2 [Biotin]](http://images.novusbio.com/fullsize2/DATA_CXCL10_DY266_ELISA_1870.jpg "N/A CXCL10/IP-10/CRG-2 [Biotin]")
![Immunocytochemistry/Immunofluorescence: HA Tag Antibody [NB600-363] - Detection of HA-tagged Tubulin in CHO cells transfected with mutant beta-tubulin cDNA encoding an HA epitope tag. Photo contributed by R. Bhattacharya and F. Cabral, University of Texas Medical School at Houston.](http://images.novusbio.com/fullsize/HA-Tag-Antibody-Immunocytochemistry-Immunofluorescence-NB600-363-img0005.jpg "Immunocytochemistry/Immunofluorescence: HA Tag Antibody [NB600-363] - Detection of HA-tagged Tubulin in CHO cells transfected with mutant beta-tubulin cDNA encoding an HA epitope tag. Photo contributed by R. Bhattacharya and F. Cabral, University of Texas Medical School at Houston.")
![Western Blot: HA Tag Antibody [NB600-363] - 200, 100, or 50 ng of E. coli whole cell lysate expressing amulti-tag fusion protein.Antibody: Affinity purified rabbitanti-HA antibody used for WB at 0.04 ug/ml(1:25,000). Detection: Chemiluminescence with anexposure time of 3 seconds.](http://images.novusbio.com/fullsize/HA-Tag-Antibody-Western-Blot-NB600-363-img0019.jpg)
![Western Blot: HA Tag Antibody [NB600-363] - Wnt/beta-catenin signaling regulates HySp5 expression. Immunoprecipitation (IP) of HA-tagged HySp5-420 expressed in HEK293T cells together or not with hudeltabeta-Catenin (upper) or huTCF1 (lower). IP was performed with an anti-HA antibody and Co-IP products were detected with the anti-beta-catenin or anti-TCF1 antibodies. Same results were obtained in two independent experiments. Each data point represents one biological independent experiment. Statistical p values: *p <= 0.05, **p <= 0.01, ***p <= 0.001, ****p <= 0.0001 (unpaired t test). Error bars indicate SD. PP primer pair, RLA relative luciferase activity Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-018-08242-2), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/HA-Tag-Antibody-Western-Blot-NB600-363-img0018.jpg)
![Western Blot: HA Tag Antibody [NB600-363] - ZFN expression levels. After co-transfection of HEK293T cells with ZFN expression vectors and pEGFP, cell lysates were probed with antibodies against the HA tag and EGFP. Amount of transfected ZFN plasmids was 75 ng, 300 ng, and 1200 ng. NT, non-transfected cells. Selection-independent generation of gene knockout mouse embryonic stem cells using zinc-finger nucleases. PLoS One (2011)](http://images.novusbio.com/fullsize/HA-Tag-Antibody-Western-Blot-NB600-363-img0017.jpg)
![Immunocytochemistry/Immunofluorescence: HA Tag Antibody [NB600-363] - HA-tagged proteins were detected in immersion fixed HEK293 human embryonic kidney cell line transfected with HA-tagged MGAT2 using 1 ug/mL goat anti-HA Tag polyclonal (NB600-362, Novus Biologicals), 1 ug/mL rabbit anti-HA Tag polyclonal (NB600-363, Novus Biologicals). Cells were stained using the appropriate secondary antibody, donkey anti-mouse IgG-NL557 (NL007) and counterstained with DAPI (blue).](http://images.novusbio.com/fullsize/HA-Tag-Antibody-Immunocytochemistry-Immunofluorescence-NB600-363-img0013.jpg)
![Western Blot: HA Tag Antibody [NB600-363] - Gene knockout in U2OS.693 cells.(A) Schematic of ZFN-mediated knockout. A ZFN pair (ZFNR & ZFNL) designed to target position 502 in the EGFP gene (E502) creates a DNA double-strand break that is sealed by the error-prone non-homologous end-joining (NHEJ) pathway & hence leads to disruption of the coding sequence. (B) Dose-dependent gene disruption in U2OS.693 cells. U2OS.693 cells that stably express a destabilized EGFP were transfected with increasing amounts of E502-specific ZFN expression vectors (75–1200 ng). The percentage of EGFP-negative cells was determined 6 days post-transfection by flow cytometry (n = 3; indicated is average & standard deviation). E502-WT, ZFN with wild-type FokI domain; E502-OH, ZFN with obligate heterodimeric FokI domain. (C) ZFN expression levels. After co-transfection of HEK293T cells with ZFN expression vectors & pEGFP, cell lysates were probed with antibodies against the HA tag & EGFP. Amount of transfected ZFN plasmids was 75 ng, 300 ng, & 1200 ng. NT, non-transfected cells. (D) Kinetics of EGFP knockout. The graph displays the percentage of EGFP-negative cells (see B) from day 1 to day 6 post-transfection for two vector amounts (n = 3; indicated is average & standard deviation). WT, E502-WT; OH, E502-OH. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/22194948), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb600-363_rabbit-polyclonal-ha-tag-antibody-310202416171456.jpg)
![Western Blot: VEGF Antibody (VG1) - BSA Free [NB100-664] - Regorafenib reduced TNBC cell migration & targeted p-STAT3/VEGF-A signaling.(A) Chemical structure of regorafenib. (B,C), Transwell assay (B) & wound-healing assay (C) were performed in TNBC cell lines after regorafenib treatment for 24 h. *p 160) or low VEGF-A (H score < = 160) levels for the indicated time of follow up. Chi-square test indicated a significant difference between VEGF-A high (N = 21) & low (N = 76) patients. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27364975), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-664_mouse-monoclonal-vegf-antibody-vg1-3102024155374.jpg)
![Western Blot: VEGF Antibody (VG1) - BSA Free [NB100-664] - Regorafenib transcriptionally inhibited VEGF-A expression through decreasing binding of STAT3 on promoter of VEGF-A.(A) Top, MDA-MB-231 cells co-transfected w/ a Renilla control vector & plasmids containing firefly luciferase gene driven by wild-type or STAT3 binding site-mutated VEGF-A promoter. After transfection for 48 h, cells treated w/ regorafenib for 24 h. Promoter activity analyzed by luciferase assay after regorafenib treatment. Bottom, MDA-MB-231 cells transfected w/ vector-control or STAT3-overexpression plasmid for 48 h. After , cells further co-transfected w/ Renilla & wild-type VEGF-A promoter & detect promoter activity as mentioned above. *p < 0.05, **p < 0.01. (B) After regorafenib treatment for 24 h, STAT3 binding site fragment detected by PCR in ChIP samples precipitated w/ STAT3 & rabbit IgG control antibodies in MDA-MB-231 cells. (C,D), MDA-MB-231 cells transfected, respectively, w/ control vector or VEGF-A overexpression plasmid (C) or STAT3 overexpression plasmid (D) for 48 h. After transfection, cells treated w/wo regorafenib for 24 h & subjected to WB assay or seeded to Transwell to analyze migration ability. *p < 0.05, **p < 0.01. (E) Cells treated w/ regorafenib at indicated dosages for 24 h & cell lysates analyzed by SHP-1 phosphatase activity assay. *p < 0.05, **p < 0.01. (F) MDA-MB-231 cells pretreated w/ pan-phosphatase inhibitor (left), or specific SHP-1 inhibitor (Middle) for 1 h before regorafenib treatment. Right, MDA-MB-231 cells transfected, respectively, w/ control siRNA or SHP-1 siRNA for 48 h. After transfection, cells treated w/wo regorafenib (10 μM) for 24 h. The protein levels analyzed by WB assay or cells seeded to Transwell to analyze migration ability. *p < 0.05, **p < 0.01. Image collected & cropped by CiteAb from following publication (//pubmed.ncbi.nlm.nih.gov/27364975), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-664_mouse-monoclonal-vegf-antibody-vg1-31020241682159.jpg)
![Analysis of a FFPE tissue section of human liver using 1:200 dilution of VEGF antibody [VG1]. The staining was developed using HRP labeled anti-rabbit secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin.](http://images.novusbio.com/fullsize/antibody/nb100-664_mouse-monoclonal-vegf-antibody-vg1-immunohistochemistry-paraffin-1211202510571.jpg)
![Analysis of a FFPE tissue section of mouse liver using 1:200 dilution of VEGF antibody [VG1]. The staining was developed using HRP labeled anti-rabbit secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin.](http://images.novusbio.com/fullsize/antibody/nb100-664_mouse-monoclonal-vegf-antibody-vg1-immunohistochemistry-paraffin-12112025105832.jpg)
![N/A FGF basic/FGF2/bFGF [HRP]](http://images.novusbio.com/fullsize2/DATA_FGF_basic_DFB00_ELISA_668.jpg "N/A FGF basic/FGF2/bFGF [HRP]")
![N/A FGF basic/FGF2/bFGF [HRP]](http://images.novusbio.com/fullsize2/FGF_basic_DFB00_ELISA_386.jpg)
![N/A TNF-alpha [Biotin]](http://images.novusbio.com/fullsize2/DATA_TNFalpha_DY510_ELISA_2079.jpg "N/A TNF-alpha [Biotin]")
![N/A cAMP [Biotin]](http://images.novusbio.com/fullsize2/DATA_cAMP_KGE002_ELISA_902.jpg "N/A cAMP [Biotin]")
![N/A cAMP [Biotin]](http://images.novusbio.com/fullsize2/DATA_cAMP_KGE002_ELISA_903.jpg)
![N/A cAMP [Biotin]](http://images.novusbio.com/fullsize2/cAMP_KGE002_ELISA_405.jpg)
![Western Blot: beta-Actin Antibody (8H10D10) [NBP1-47423] - Analysis using anti-Beta Actin mAb against human, mouse, hamster, rat, and primate cell lystate(s): (1) NIH/3T3, (2) Jurkat, (3) HeLa, (4) CHO, (5) PC12, (6) HEK293, (7) COS, (8) A549, and (9) MCF-7. A specific band can be observed at a molecular weight of approximately 43 kDa in all lanes.](http://images.novusbio.com/fullsize/beta-Actin-Antibody-8H10D10-Western-Blot-NBP1-47423-img0006.jpg "Western Blot: beta-Actin Antibody (8H10D10) [NBP1-47423] - Analysis using anti-Beta Actin mAb against human, mouse, hamster, rat, and primate cell lystate(s): (1) NIH/3T3, (2) Jurkat, (3) HeLa, (4) CHO, (5) PC12, (6) HEK293, (7) COS, (8) A549, and (9) MCF-7. A specific band can be observed at a molecular weight of approximately 43 kDa in all lanes.")
![Immunocytochemistry/Immunofluorescence: beta-Actin Antibody (8H10D10) [NBP1-47423] - Beta actin was detected in NIH-3T3 cells fixed with methanol using mouse anti-mouse beta-Actin monoclonal antibody (NBP1-47423) at 1:5400 dilution. Cells were stained using NorthernLights 557 conjugated anti-mouse secondary antibody (NL007) and counterstained with DAPI.](http://images.novusbio.com/fullsize/beta-Actin-Antibody-8H10D10-Immunocytochemistry-Immunofluorescence-NBP1-47423-img0011.jpg)
![Simple Western: beta-Actin Antibody (8H10D10) [NBP1-47423] - Image shows a specific band for Beta Actin in 1.0 mg/mL of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.](http://images.novusbio.com/fullsize/beta-Actin-Antibody-8H10D10-Simple-Western-NBP1-47423-img0009.jpg)
![Flow Cytometry: beta-Actin Antibody (8H10D10) [NBP1-47423] - Analysis of HeLa cells using mouse Monoclonal beta-Actin antibody (Orange) and Isotype control Antibody (Blue).](http://images.novusbio.com/fullsize/beta-Actin-Antibody-8H10D10-Flow-Cytometry-NBP1-47423-img0010.jpg)
![Western Blot: beta-Actin Antibody (8H10D10) [NBP1-47423] - TTF-1 alters the phosphorylation of GSK3alpha/beta. (A) Panels of immunoblot images examining total GSK3alpha/beta and phosphorylation at specific Ser or Tyr residues in A549 transfectant cells. Beta-Actin is included as a loading control. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-019-44549-w), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/beta-Actin-Antibody-8H10D10-Western-Blot-NBP1-47423-img0014.jpg)
![Immunocytochemistry/Immunofluorescence: beta-Actin Antibody (8H10D10) [NBP1-47423] - HeLa cells were fixed and permeabilized for 10 minutes with -20C MeOH. The cells were incubated with beta-Actin Antibody [8H10D10] conjugated to Biotin (NBP1-47423B) at 5ug/ml overnight at 4C and detected with Streptavidin conjugated to DyLight 550. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.](http://images.novusbio.com/fullsize/beta-Actin-Antibody-8H10D10-Immunocytochemistry-Immunofluorescence-NBP1-47423-img0015.jpg)
![Flow Cytometry: beta-Actin Antibody (8H10D10) - BSA Free [NBP1-47423] - An intracellular stain was performed on NIH3T3 cells with beta-Actin Antibody (8H10D10) NBP1-47423 (blue) and a matched isotype control MAB004 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (84540, Thermo Fisher).](http://images.novusbio.com/fullsize/beta-Actin-Antibody-8H10D10-BSA-Free-Flow-Cytometry-NBP1-47423-img0017.jpg)
![Western Blot: beta-Actin Antibody (8H10D10) [NBP1-47423] - Western blot of pancreatic cell lines (CAPAN2, PATU8988T, DANG, and PANC1005). Primary antibody: beta-Actin (8H10D10), NBP1-47423, 1:1000 dilution. Secondary antibody: HRP conjugated Mouse IgG (H+L), NB7539. A specific band can be observed at a molecular weight of approximately 42 kDa. Image from verified customer review.](http://images.novusbio.com/fullsize/beta-Actin-Antibody-8H10D10-Western-Blot-NBP1-47423-img0007.jpg)
![Western Blot: beta-Actin Antibody (8H10D10) [NBP1-47423] - Analysis of Beta Actin expression in 1) HeLa 2) HepG2 and 3) Cos7 whole cell lysates.](http://images.novusbio.com/fullsize/beta-Actin-Antibody-8H10D10-Western-Blot-NBP1-47423-img0008.jpg)
![Western Blot: beta-Actin Antibody (8H10D10) [NBP1-47423] - Lysates of HeLa human cervical epithelial carcinoma cell line, A431 human epithelial carcinoma cell line, K562 human chronic myelogenous leukemia cell line, NIH-3T3 mouse embryonic fibroblast cell line, and C6 rat glioma cell line were probed with 1:2500 mouse anti-beta-Actin monoclonal (NBP1-47423) followed by 1:2000 dilution of donkey anti-mouse IgG-HRP secondary antibody (HAF018).](http://images.novusbio.com/fullsize/beta-Actin-Antibody-8H10D10-Western-Blot-NBP1-47423-img0012.jpg)
![Western Blot: beta-Actin Antibody (8H10D10) [NBP1-47423] - Three different human cancer cell lines (SW780, BT-20, and CLS439) were probed with the antibody. Image from verified customer review.](http://images.novusbio.com/fullsize/beta-Actin-Antibody-8H10D10-Western-Blot-NBP1-47423-img0013.jpg)
![Flow Cytometry: beta-Actin Antibody (8H10D10) [NBP1-47423] - Analysis of MCF-7 cells using anti-Beta Actin mAb (right) and negative control (left).](http://images.novusbio.com/fullsize/beta-Actin-Antibody-8H10D10-Flow-Cytometry-NBP1-47423-img0004.jpg)
![Flow Cytometry: beta-Actin Antibody (8H10D10) - BSA Free [NBP1-47423] - An intracellular stain was performed on U-251 MG cells with beta-Actin Antibody (8H10D10) NBP1-47423 (blue) and a matched isotype control MAB004 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (84540, Thermo Fisher).](http://images.novusbio.com/fullsize/beta-Actin-Antibody-8H10D10-BSA-Free-Flow-Cytometry-NBP1-47423-img0016.jpg)
![N/A GDF-15 [HRP]](http://images.novusbio.com/fullsize2/GDF15_DGD150_ELISA_2662.jpg "N/A GDF-15 [HRP]")
![N/A GDF-15 [HRP]](http://images.novusbio.com/fullsize2/GDF15_DGD150_ELISA_2661.jpg)
![N/A GDF-15 [HRP]](http://images.novusbio.com/fullsize2/GDF15_DGD150_ELISA_698.jpg)
![N/A GDF-15 [HRP]](http://images.novusbio.com/fullsize2/GDF15_DGD150_ELISA_108.jpg)
![Immunocytochemistry/Immunofluorescence: GFP Antibody [NB100-1614] - Staining of cultured hippocampal neuron from rat. ICC/IF image submitted by a verified customer review.](http://images.novusbio.com/fullsize/GFP-Antibody-Immunocytochemistry-Immunofluorescence-NB100-1614-img0003.jpg "Immunocytochemistry/Immunofluorescence: GFP Antibody [NB100-1614] - Staining of cultured hippocampal neuron from rat. ICC/IF image submitted by a verified customer review.")
![Immunocytochemistry/Immunofluorescence: GFP Antibody [NB100-1614] - HeLa cells transiently expressing GFP were visualized using GFP antibody (green). Nuclei were counterstained with DAPI. ICC/IF image submitted by a verified customer review.](http://images.novusbio.com/fullsize/GFP-Antibody-Chromatin-Immunoprecipitation-NB100-1614-img0007.jpg)
![Immunocytochemistry/Immunofluorescence: GFP Antibody [NB100-1614] - IF analysis of GFP in mouse trigeminal ganglia, cornea. Image courtesy of product review by Jessica Newsom.](http://images.novusbio.com/fullsize/GFP-Antibody-Immunocytochemistry-Immunofluorescence-NB100-1614-img0001.jpg)
![Immunohistochemistry: GFP Antibody [NB100-1614] - 5dpf Tg(brn3c:GFP) larvae were incubated with vehicle alone (DMSO, A), 400uM of cisplatin (CP, B), pre-treated with Qx for 2 hours and then incubated with CP for 6 hours (Qx, CP, C,E,G) or pre-treated with Qx for 2 hours and then co-treated with Qx and CP for 6 more hours (Qx+CP, D,F,H). Animals were fixed and immunostained for GFP (green) and otoferlin (red). Image collected and cropped by CiteAb from the following publication (nature.com/articles/s41598-018-33520-w), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/GFP-Antibody-Immunohistochemistry-NB100-1614-img0018.jpg)
![Immunocytochemistry/Immunofluorescence: GFP Antibody [NB100-1614] - IF analysis of GFP in rat cortical culture (2 weeks old). ICC/IF image submitted by a verified customer review.](http://images.novusbio.com/fullsize/GFP-Antibody-Chromatin-Immunoprecipitation-NB100-1614-img0008.jpg)
![Immunocytochemistry/Immunofluorescence: GFP Antibody [NB100-1614] - LNCaP human prostate cancer cell line imaged after transfection with GFP expressing plasmid. ICC/IF image submitted by a verified customer review.](http://images.novusbio.com/fullsize/GFP-Antibody-Immunocytochemistry-NB100-1614-img0009.jpg)
![Immunocytochemistry/Immunofluorescence: GFP Antibody [NB100-1614] - Staining of a tissue section through the midbrain region of an adult mouse for GFP (green) and Tryptophan Hydroxylase-positive neurons (RED).](http://images.novusbio.com/fullsize/GFP-Antibody-Immunocytochemistry-Immunofluorescence-NB100-1614-img0010.jpg)
![Immunohistochemistry-Paraffin: GFP Antibody [NB100-1614] - Pyramidal neurons in the hippocampal formation of a neonatal mouse brain. Tissue was paraformaldehyde-fixed (4%) and paraffin-embedded. GFP staining is in green in left panel.](http://images.novusbio.com/fullsize/GFP-Antibody-Immunohistochemistry-Paraffin-NB100-1614-img0011.jpg)
![Immunohistochemistry-Paraffin: GFP Antibody [NB100-1614] - Embyonic mouse brain section. GFP in green, DAPI in grey. IHC-P image submitted by a verified customer review.](http://images.novusbio.com/fullsize/GFP-Antibody-Immunohistochemistry-Paraffin-NB100-1614-img0016.jpg)
![Immunohistochemistry: GFP Antibody [NB100-1614] - (J) control. (K) Neo 200uM 30min. (L) GM 50uM 1 hour. (M) CP: 400uM 2 hours. (N) Qx 300uM 8 hours. (O) Qx 300uM 8 hours + Neo 200uM 30min. (P) Qx 300uM 8 hours + GM 50uM 1 hour. (Q) Qx 300uM 8 hours + CP 400uM 2 hours. Asterisks denote neuromast supporting cells positive for BrdU. Image collected and cropped by CiteAb from the following publication (nature.com/articles/s41598-018-33520-w), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/GFP-Antibody-Immunohistochemistry-NB100-1614-img0017.jpg)
![Immunohistochemistry: Chicken Polyclonal GFP Antibody [NB100-1614] - Chicken Polyclonal GFP Antibody on mouse cancer tissue. DAPI (Red) and Cytopastmic GFP stain (Green). Primary antibody dilution: 1:200 in a 200 um slice. Image from a verified customer review.](http://images.novusbio.com/fullsize/antibody/nb100-1614_chicken-polyclonal-gfp-antibody-immunohistochemistry-1610202310250..png)
![Immunohistochemistry-Frozen: Chicken Polyclonal GFP Antibody [NB100-1614] - Image depicting GFP in magenta and CD31 (Catalog # AF3628) in red. Tissue is a fixed frozen section of sciatic nerve obtained from a S100A4 GFP mouse. NB100-1614 was diluted 1 in 10000 and was left on tissue sections overnight at 4 degrees Celsius. Secondary antibody was donkey anti-chicken conjugated to Alexa 647. Image from a verified customer review.](http://images.novusbio.com/fullsize/antibody/nb100-1614_chicken-polyclonal-gfp-antibody-immunohistochemistry-frozen-842024125355..jpg)
![Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB100-1614] - Macrophages regulate dormancy in tumor cells.a Representative image of triple immunofluorescently stained in E0771-GFP primary tumor tissue for tumor cells, macrophages, & NR2F1. Green = GFP; Red = NR2F1; White = IBA-1; Blue = DAPI. White arrow shows a macrophage. The yellow arrow contact between an NR2F1-positive tumor cell & a macrophage. Mϕ=Macrophage. Scale bar=20 μm. b Quantification showing frequency of distances between NR2F1+ tumor cells to nearest macrophage in primary tumor. Data is normalized to frequency of distances between all DAPI+ nuclei to nearest TMEM. Bar = mean. Error bars = ±SEM. n = 34 fields of view (551 × 316 µm2) in 4 animals. For comparison between 0 & 200 µm bins a two-tailed Mann-Whitney test used (p < 0.0001). ****p < 0.0001. c Representative immunofluorescence images of NR2F1 expression in E0771-GFP tumor cells cultured alone, in direct contact w/ BAC1.2F5 macrophages,/in direct contact w/ HUVEC endothelial cells. White arrows show macrophages/endothelial cells in direct contact w/ a tumor cell. Green = GFP; Red = NR2F1; Blue = DAPI. TC = Tumor Cell. Mϕ = Macrophage. EC = Endothelial Cell. Scale bar = 15 μm. d Percentage of NR2F1-positive tumor cells from each group in C. TC alone: n = 777 cells in 9 independent experiments; TC+Mϕ; n = 226 cells in 6 independent experiments, TC+EC = n = 359 cells in 4 independent experiments. Bar = mean. Error bars = ±SEM. For TC vs. TC+Mϕ (p = 0.0039), & for TC vs. TC+EC (p = 1), a two-tailed Kruskal-Wallis test w/ Dunn’s multiple comparisons adjustment used. For TC+Mϕ vs. TC+EC (0.012), a two-tailed one-way ANOVA w/ Sidak’s multiple comparison adjustment used. *p < 0.05. **p < 0.01; ns = not significant. Source data are provided as a Source Data file. Image collected & cropped by CiteAb from following publication (//pubmed.ncbi.nlm.nih.gov/35110548), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-1614_chicken-polyclonal-gfp-antibody-31020241554535.jpg)
![Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB100-1614] - Spontaneously metastasizing tumor cells survive significantly longer at the secondary site compared to intravenously injected tumor cells.a Representative intravital microscopy images showing the possible fates of extravascular disseminated tumor cells in the lung parenchyma. Top: Images of disseminated tumor cells just after extravasation. Bottom left: Example of an extravascular tumor cell, which has died, as evidenced by small extravascular apoptotic bodies (yellow arrow). Bottom middle: Example of an extravascular tumor cell that survived as a single & solitary tumor cell over time. Bottom right: Example of an extravascular tumor cell that began to divide & grow into a micro-metastasis. Red = tdTomato labeled endothelial cells & 155 kDa Tetramethylrhodamine dextran labeled blood serum, Green = GFP labeled tumor cells. Yellow dashed lines delineate blood vessel boundaries. Scale bar = 15 μm. b Percentage of extravascular E0771-GFP disseminated tumor cells that died, survived, or grew after extravasation in EM & SM models 64 hrs after arrival to the lung vasculature. EM: n = 27 tumor cells in 4 mice. SM: n = 31 tumor cells in 4 mice. Bar = mean. Error bars = ±SEM. For Died & Survived columns, a two-tailed unpaired t-test was used (p = 0.0003 & 0.0005, respectively). For Grew columns, a two-tailed Mann-Whitney test was used (p = 0.14). ***p < 0.001. ns = not significant. Source data are provided as a Source Data file. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35110548), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-1614_chicken-polyclonal-gfp-antibody-31020241553510.jpg)
![Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB100-1614] - Qx protects against ototoxin-induced HC death & promotes supporting cell proliferation. (A–I) TUNEL assay (red) performed in zebrafish incubated w/ vehicle or 300 µM of Qx alone (controls) or w/ corresponding ototoxin w/ or w/out Qx 300 µM. Animals counterstained w/ phalloidin (green). (A) Neomycin (Neo) 200 µM incubation for 30 min. (B) Gentamicin (GM) 50 µM incubation for 1 hour. (C) Cisplatin (CP) 400 µM incubation for 2 hrs. (D) GM 50 µM incubation for 1 hour followed by recovery for 5 hrs. (E) Incubation w/ Qx 300 µM for 8 hrs & Neo 200 µM for 30 min. (F) Incubation w/ Qx 300 µM for 8 hrs & GM 50 µM for 1 hour. (G) Qx 300 µM for 8 hrs + CP 400 µM for 2 hrs. (H) Qx 300 µM incubation for 8 hrs + GM 50 µM for 1 hour followed by 5 hrs recovery. Asterisks denote TUNEL-positive HCs. (I) The % of TUNEL-positive neuromasts calculated for each treatment & represented as mean +/− SEM. (J–R) Proliferation assays performed in 5dpf Tg(brn3c:GFP) in presence/absence of Qx & corresponding ototoxin, by BrdU-labelling method (red). Animals immunostained for GFP (green). (J) control. (K) Neo 200 µM 30 min. (L) GM 50 µM 1 hour. (M) CP: 400 µM 2 hrs. (N) Qx 300 µM 8 hrs. (O) Qx 300 µM 8 hrs + Neo 200 µM 30 min. (P) Qx 300 µM 8 hrs + GM 50 µM 1 hour. (Q) Qx 300 µM 8 hrs + CP 400 µM 2 hrs. Asterisks denote neuromast supporting cells positive for BrdU. (R) The % of BrdU-positive supporting cells per neuromast calculated for each treatment & represented as mean +/− SEM. One-way ANOVA, *p < 0.05, **p < 0.01. Black asterisks compared versus corresponding control. Red asterisk compared versus corresponding ototoxin-only treatment. Scale bar: (A–H) 10 µm, (J–O) 7 μm. Data taken from at least 15 animals & 3 experiments runs. Image collected & cropped by CiteAb from following publication (//pubmed.ncbi.nlm.nih.gov/30310154), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-1614_chicken-polyclonal-gfp-antibody-3102024155524.jpg)
![Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB100-1614] - Spontaneously metastasizing tumor cells are more frequently doubly positive for dormancy & stem-like markers compared to intravenously injected tumor cells.a Representative images of triple immunofluorescence staining for GFP, NR2F1, & SOX9 expression in primary tumors, circulating tumor cells (CTCs), & disseminated tumor cells (Lung) from an E0771-GFP SM model (Left) & in disseminated tumor cells (Lung) from an EM model (Right). Green = GFP; Red = NR2F1; Orange = SOX9; Blue = DAPI. Scale bar for Primary Tumor = 50 μm. Scale bar for CTCs & Lung = 15 μm. b Percentage of double-positive tumor cells NR2F1-positive SOX9High from each group in Fig. 5a. Primary Tumor: n = 2383 in 97 fields of view (65 × 65 µm2) in 7 animals; CTCs: n = 379 cells in 8 animals; SM Lung: n = 104 cells in 9 animals; In vitro: n = 413 cells in 3 independent experiments. EM Lung: n = 75 cells in 7 animals. Bar = mean. Error bars = ±SEM. For EM Lung vs. SM Lung (p = 0.0001) & EM Lung vs. in vitro (p = 0.69), a two-tailed Kruskal-Wallis test with Dunn’s multiple comparisons adjustment was used. For PT vs. CTC: (p = 0.0041), PT vs. Lung SM (p = 0.0030), & CTC vs. Lung SM (p = 1.00) a two-tailed ANOVA test with Sidak’s multiple comparisons adjustment was used. **p < 0.01. ***p < 0.001. ns = not significant. Source data are provided as a Source Data file. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35110548), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-1614_chicken-polyclonal-gfp-antibody-310202416571.jpg)
![Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB100-1614] - Dose protection curve against aminoglycosides. 5dpf larvae were incubated with vehicle (E3, A) or with 50 μM (D–F), 150 μM (G–I) or 300 μM (J–L) of Qx for a total of 8 hours. Gentamicin (GM, 50 μM, B,E,H,K) or neomycin (Neo, 200 µM,C,F,I,L) were added during the last 60 min or 30 min of incubation, respectively. Animals were fixed & stained for otoferlin (red) & GFP (green). (M) Quantification of the number of hair cells per neuromast after the different treatments represented as mean +/− SEM. Note that since no significant differences were found in the number of hair cells per neuromast when animals were incubated with the different Qx concentrations (0–300 µM, A,D,G,J), the control value represents the average of all these treatments. One-way ANOVA, Dunnett post test. *p < 0.05, **p < 0.01, ***p < 0.001. Black asterisks compared versus control. Red asterisks compared versus the corresponding aminoglycoside-only treatment. (N) Scores for neuromast morphology (see Materials & Methods). Scale bar: 7 μm. Data were taken from at least 20 animals & 3 experiments runs. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30310154), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-1614_chicken-polyclonal-gfp-antibody-31020241555229.jpg)
![Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB100-1614] - Qx protects against cisplatin ototoxicity. 5dpf Tg(brn3c:GFP) larvae were incubated with vehicle alone (DMSO, A), 400 μM of cisplatin (CP, B), pre-treated with Qx for 2 hours & then incubated with CP for 6 hours (Qx, CP, C,E,G) or pre-treated with Qx for 2 hours & then co-treated with Qx & CP for 6 more hours (Qx + CP, D,F,H). Animals were fixed & immunostained for GFP (green) & otoferlin (red). (I) Quantification of the number of hair cells per neuromast after the different treatments represented as mean +/− SEM. One-way ANOVA, Dunnett post test. *p < 0.05, ***p < 0.001. Black asterisks compared versus control. Red asterisks compared versus CP 400 µM. (J) Scores for neuromast morphology (see Materials & Methods). Scale bar: 6 μm. Data were taken from at least 20 animals & 3 experiments runs. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30310154), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-1614_chicken-polyclonal-gfp-antibody-31020241612686.jpg)
![Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB100-1614] - Dose protection curve against CP. 5dpf Tg(brn3c:GFP) larvae were incubated with 50 µM to 800 µM of CP (A,C,E,G) for 6 hours or pre-treated with 300 µM of Qx for 2 hours & then co-treated with Qx & CP (50µM-800µM) for 6 hours (B,D,F,H). Animals were fixed & immunostained for GFP (green) & otoferlin (red). Control animals were exposed to vehicle alone (DMSO). (I) Quantification of the number of hair cells per neuromast after the different treatments represented as mean +/− SEM. One-way ANOVA, Dunnett post test. ***p < 0.001. Black asterisks compared versus DMSO-treated animals. Red asterisks compared versus the corresponding CP concentration. (J) Scores for neuromast morphology (see Materials & Methods). Scale bar: 6 μm. Data were taken from at least 20 animals & 3 experiments runs. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30310154), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-1614_chicken-polyclonal-gfp-antibody-310202415541991.jpg)
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