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NB100-2220

LC3B Antibody

(39 Reviews)

(814 Publications)

0.1 ml

NB100-2220

Knockout Validated: LC3B Antibody [NB100-2220] - Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and LC3B knockout HeLa cell line (KO) untreated (-) or treated (+) with 50 uM Chloroquine for 18 hours. PVDF membrane was probed with 0.5 ug/mL of Rabbit Anti-LC3B Polyclonal Antibody (Catalog # NB100-2220) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog# HAF008). A specific band was detected for LC3B at approximately 15 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. GAPDH is shown as a loading control. This experiment was conducted under reducing conditions.

<b>Biological Strategies Validation and Genetic Strategies Validation.</b>Western Blot: LC3B Antibody [NB100-2220] - Detection of LC3B in mouse ES cell lysates. Atg5-/- ES cells from Dr. Noboru Mizushima [Mizushima, N. et al. J. Cell Biol. 152 (2001)] Photo courtesy of Dr. Beth Levine, UT SW Medical Center.

<b>Biological Strategies Validation. </b>Western Blot: LC3B Antibody [NB100-2220] - Western blot analysis of HeLa (1), HeLa + CQ (2), SHSY5Y (3), SHSY5Y +CQ (4), A431 (5), A431 +CQ (6) and Ntera2 (7) using LC3 antibody at 2 ug/ml.

<b>Genetic Strategies Validation. </b>Knockout Validated: LC3B Antibody [NB100-2220] - LC3B was detected in immersion fixed Cloroquine treated Hela cells (left) but was not detected in LC3B knockout Hela cells (right) using rabbit anti-human LC3B polyclonal antibody (Catalog #NB100-2220) at 0.3 ug/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm.

<b>Biological Strategies Validation. </b>Western Blot: LC3B Antibody [NB100-2220] - shows lysates of mouse NIH3T3 and rat PC-12 cell lines untreated (-) or treated (+) with Chloroquine. PVDF membrane was probed with 0.5 ug/mL rabbit anti-LC3B polyclonal Antibody (NB100-2220, Novus Biologicals), followed by 1:2000 dilution of goat anti-rabbit IgG secondary antibody.

<b>Biological Strategies Validation. </b>Western Blot: LC3B Antibody [NB100-2220] - Analysis of LC3B in Huh-7 and SMMC-7721 cells using anti-LC3B antibody. Image from verified customer review.

Western Blot: LC3B Antibody [NB100-2220] - Analysis using the Biotin conjugate of NB100-2220. Image from verified customer review.

Immunohistochemistry-Paraffin: LC3B Antibody [NB100-2220] - Human ovarian Cancer tissue stained using heat mediated antigen retrieval in pH 6.0 citrate buffer at 1:200 dilution. Image provided by verified customer review.

Immunocytochemistry/Immunofluorescence: LC3B Antibody [NB100-2220] - LC3B detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with Chloroquine using 1ug/mL rabbit anti-LC3B polyclonal (NB100-2220, Novus Biologicals). Cells were stained using donkey anti-rabbit IgG-NL557 and counterstained with DAPI (blue).

Western Blot: LC3B Antibody [NB100-2220] - Autophagy Investigation. (A) Representative WBs of LC3B-I, LC3B-II and GAPDH loading control in DANs derived from 4 iPD patients and 4 healthy controls. (B) WBs from three independent experiments were quantified by densitometry and LC3B-II normalized to a loading control. Grouping of data by healthy vs iPD as well as iPD stratified for T/T and C/C genotype. (C) Ratio of LC3B-II (normalized to loading control) following bafilomycin treatment/untreated. Citation: Marrone L, Bus C, Schondorf D, Fitzgerald JC, Kubler M, Schmid B, et al. (2018) Generation of iPSCs carrying a common LRRK2 risk allele for in vitro modeling of idiopathic Parkinson's disease. PLoS ONE 13(3): e0192497. //doi.org/10.1371/journal.pone.0192497

Immunocytochemistry/Immunofluorescence: LC3B Antibody [NB100-2220] - LC3B antibody was tested in HeLa cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red).

Immunohistochemistry: LC3B Antibody [NB100-2220] - Analysis using the Biotin conjugate of NB100-2220. Staining of brain, cerebral cortex, neurons with cell processes.

<b>Biological Strategies Validation. </b>Western Blot: LC3B Antibody [NB100-2220] - Analysis of LC3 in H9C2 cells.

Western Blot: LC3B Antibody [NB100-2220] - Analysis using the DyLight 550 conjugate of NB100-2220. Detection of LC3 in pancreatic cancer cells.

Western Blot: LC3B Antibody [NB100-2220] - Detection of LC3B in mouse HL-1 cell lysate. Photo courtesy of product review by verified customer.

<b>Biological Strategies Validation. </b>Western Blot: LC3B Antibody [NB100-2220] - WB detection of LC3I and LC3II in mouse cochlea cell line SV-K1. Cells were treated with chloroquine (1uM), and As2O3 (1uM) for 24 hrs. This image was submitted via customer review.

Western Blot: LC3B Antibody [NB100-2220] - Detection in fibroblast whole cell extract. Image from verified customer review.

Western Blot: LC3B Antibody [NB100-2220] - Macrophage lysates probed with anti-LC3B antibody. This image was submitted via customer Review.

<b>Biological Strategies Validation. </b>Western Blot: LC3B Antibody [NB100-2220] - Uncropped western blots used for quantification. Detection of LC3B. Normalized Protein levels by the indicated housekeeping protein. iPD genotypes are reported. Blots show protein levels in the presence or absence of a specified treatment. Baf = bafilomycin, Rot = rotenone, Val = valinomycin, CZC = CZC-25146Citation: Marrone L, Bus C, Schondorf D, Fitzgerald JC, Kubler M, Schmid B, et al. (2018) Generation of iPSCs carrying a common LRRK2 risk allele for in vitro modeling of idiopathic Parkinson's disease. PLoS ONE 13(3): e0192497. //doi.org/10.1371/journal.pone.0192497

Immunocytochemistry/Immunofluorescence: LC3B Antibody [NB100-2220] - HeLa cells were treated with 50uM CQ overnight, fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton-X100. The cells were incubated with anti-LC3B at 2 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.

Immunocytochemistry/Immunofluorescence: LC3B Antibody [NB100-2220] - Untreated HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton-X100. The cells were incubated with anti-LC3B at 2 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.

Immunohistochemistry: LC3B Antibody [NB100-2220] - IHC of rat brain from confirmed customer review.

Immunohistochemistry-Paraffin: LC3B Antibody [NB100-2220] - IHC analysis of a formalin fixed and paraffin embedded tissue section of mouse brain using 1:200 dilution of rabbit anti-LC3B antibody. The specific signal of LC3 was detected using HRP-conjugated secondary antibody with DAB reagent, and nuclei of cells were counterstained using hematoxylin. This LC3B antibody generated a low to moderate levels of cytoplasmic staining in the glial cells. The neurons depicted a moderate to strong staining for LC3 in their cytoplasm.

Immunohistochemistry-Frozen: LC3B Antibody [NB100-2220] - LC3 accumulation in muscle fibers from patients with PAD (row B - E). Normal fibers from a Non-PAD sample (row A). Scale bar = 100 uM. This image was submitted via customer Review.

Simple Western: LC3B Antibody [NB100-2220] - Simple Western lane view shows a specific band for LC3B in 0.5 mg/ml of Neuro2A lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Reactivity:

Hu, Mu, Rt, Po, Av, Ba, Bv, Ca, Pm, SyHa, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Av, Ba, Bv, Ca, Pm, SyHa, Ze

Applications:

, , , , , , , , , , ,

Applications: , , , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 405	NB100-2220AF405
Alexa Fluor 488	NB100-2220AF488
Alexa Fluor 647	NB100-2220AF647
Alexa Fluor 700	NB100-2220AF700
Biotin	NB100-2220B
DyLight 405	NB100-2220V
DyLight 488	NB100-2220G
DyLight 550	NB100-2220R
DyLight 650	NB100-2220C
FITC	NB100-2220F
HRP	NB100-2220H
Janelia Fluor 549	NB100-2220JF549
Janelia Fluor 646	NB100-2220JF646

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NB100-105

HIF-1 alpha Antibody (H1alpha67)

(33 Reviews)

(757 Publications)

0.1 ml

NB100-105

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - HIF-1 alpha induction by CoCl2 on Caki-1 cell lysate. Image from verified customer review.

<b>Biological Strategies Validation and Genetic Strategies Validation.</b>Knockout Validated: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - HIF-1 alpha was detected in immersion fixed DFO treated Hela cells (left) but was not detected in HIF-1 knockout HeLa cells (right) using Mouse Anti-human HIF-1 alpha monoclonal antibody (Catalog #NB100-105) at 25 ug/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to nuclei.

<b>Biological Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Analysis using the HRP conjugate of NB100-105. Detection of 50ug cobalt chloride induced COS-7 nuclear extracts (NB800-PC26) using NB100-105.

<b>Biological Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Analysis of HIF-1 alpha in human hepatocytes from cancer patient using anti-HIF-1 alpha antibody. Image from verified customer review.

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Staining of HIF1 alpha in human kidney. Renal tubular epithelium showed moderate membranous, cytoplasmic and nuclear staining, and glomeruli showed faint to moderate nuclear staining.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Analysis using the Biotin conjugate of NB100-105. Staining of human glioblastoma multiforme.

Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Expression of HIF-1 in intresititial pig cells under 12% of 02 condition. Image from verified customer review.

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - HIF-1a was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with DFOA using 1 ug/mL of mouse anti-HIF-1a monoclonal (NB100-105, Novus Biologicals). Cells were stained using donkey anti-rabbit and counterstained with DAPI (blue).

Flow Cytometry: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Analysis using the Alexa Fluor (R) 488 conjugate of NB100-105. Staining of HIF-1 alpha in H929 cells using HIF-1 alpha antibody. Image from verified customer review.

Flow Cytometry: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Analysis using the Alexa Fluor (R) 488 conjugate of NB100-105. Staining of HIF-1 alpha in multiple myeloma cells: H929 cells (0.5x10^6) were stained with Alexa Fluor 488 (R) conjugated HIF-1a antibody (NB100-105AF488). Image courtesy of Dr. Barbara Muz at Washington University in St. Louis School of Medicine.

<b>Biological Strategies Validation and Genetic Strategies Validation.</b>Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Figure Legend: Naive CD4 T cells from WT, VHL-deficient (Vhl KO), or HIF-1a-deficient (Hif1a KO) mice were differentiated under IL-22-skewing conditions for a total of 60 h. Some cells remained at normoxia for the duration of the culture (N); others were at normoxia for 35 h and then hypoxia (1% O2) for 24 h (H). At 60 h, nuclear extracts were harvested, and HIF-1a and Lamin B1 levels were analyzed by Western blot. This image was submitted via customer Review. (Mouse, WB)

Reactivity:

Hu, Mu, Rt, Po, Bv, Ft, Mk, Pm, Rb, Sh, Xp

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, Ft, Mk, Pm, Rb, Sh, Xp

Applications:

, , , , , , , , , , , , ,

Applications: , , , , , , , , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 405	NB100-105AF405
Alexa Fluor 488	NB100-105AF488
Alexa Fluor 647	NB100-105AF647
Alexa Fluor 700	NB100-105AF700
Allophycocyanin	NB100-105APC
Allophycocyanin/Cy7	NB100-105APCCY7
Biotin	NB100-105B
DyLight 350	NB100-105UV
DyLight 405	NB100-105V
DyLight 488	NB100-105G
DyLight 550	NB100-105R
DyLight 650	NB100-105C
DyLight 680	NB100-105FR
DyLight 755	NB100-105IR
FITC	NB100-105F
HRP	NB100-105H
Janelia Fluor 549	NB100-105JF549
Janelia Fluor 646	NB100-105JF646
PE	NB100-105PE
PE/Atto594	NB100-105PEATT594
PE/Cy5.5	NB100-105PECY55
PE/Cy7	NB100-105PECY7
PerCP	NB100-105PCP

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NB100-122

HIF-2 alpha/EPAS1 Antibody

(29 Reviews)

(562 Publications)

0.1 ml

NB100-122

Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - 786-O cells without or with VHL overexpression. Image from verified customer review.

<b>Independent Antibodies Validation, Biological Strategies Validation and Genetic Strategies Validation.</b>Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - HIF-2 alpha in MDA-MB-231 cell lysate (overexpression and endogenous samples) using anti-HIF-2 alpha antibody. HIF-2 alpha antibody did not react to HIF-1 alpha overexpression. Image from verified customer review.

<b>Biological Strategies Validation. </b>Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - HIF-2 alpha in human retinal and choroidal primary endothelia lysates using anti-HIF-2 alpha antibody. Image from verified customer review.

Immunocytochemistry/Immunofluorescence: HIF-2 alpha/EPAS1 Antibody [NB100-122] - HIF-2 alpha antibody was tested in RCC4 cells at a 1:200 dilution against DyLight 488 (Green). Alpha tubulin and nuclei were counterstained against DyLight 568 (Red) and DAPI (Blue), respectively.

<b>Biological Strategies Validation. </b>Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Analysis using the HRP conjugate of NB100-122. Detection of normoxic and hypoxic nuclear rat cell lysates.

<b>Biological Strategies Validation. </b>Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Lane 1: Cobalt chloride treated COS7 nuclear extracts. Lane 2: Untreated COS7 nuclear extracts.

Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - HIF2 expression in HIF2 over-expressed HEK 293T cells. Image from verified customer review.

Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Expression and correlation of 3 antibodies: HIF-1 alpha, HIF-2 alpha and HAF in ccRCC. (A, C & E) Positive nuclear staining to the 3 antibodies: HIF-1 alpha, HIF-2 alpha and HAF, in primary ccRCCs at high magnification (100X). (B, D & F) Heterogenous nuclear staining of 3 antibodies in a tumor at low magnification (20X). (G, H and I). Correlation of 3 antibodies with the Fuhrman grade.Citation: Ambrosetti D, Dufies M, Dadone B, Durand M, Borchiellini D, Amiel J, et al. (2018) The two glycolytic markers GLUT1 and MCT1 correlate with tumor grade and survival in clear-cell renal cell carcinoma. PLoS ONE 13(2): e0193477. //doi.org/10.1371/journal.pone.0193477

Chromatin Immunoprecipitation: HIF-2 alpha/EPAS1 Antibody [NB100-122] - MCF7 cells were exposed to 20% or 1% O2 for 16h, and ChIP assays were performed using IgG or antibodies against HIF-1a, HIF-2a, HIF-1b. Image from verified customer review.

<b>Biological Strategies Validation. </b>Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Analysis on normoxic and hypoxic nuclear rat cell lysates.

Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Human Glioma whole cell lysates. Image from verified customer review.

Immunohistochemistry-Paraffin: HIF-2 alpha/EPAS1 Antibody [NB100-122] - HIF-2 alpha immunoreactivity in human cardiac myocytes stained with NB100-122.

Simple Western: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Image shows a specific band for HIF-2 alpha in 0.5 mg/mL of hypoxic HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Immunoblotting: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Analysis of protein extracts by immunoblotting against the N-terminus of VDAC1 isoform (VDAC1-Nter), the three VDAC isoforms (VDACs poly), ISCU and CAIX/ HIF-2 alpha/EPAS1 antibody. Citation: Ferecatu I, Canal F, Fabbri L, Mazure NM, Bouton C, Golinelli-Cohen M-P (2018) Dysfunction in the mitochondrial Fe-S assembly machinery leads to formation of the chemoresistant truncated VDAC1 isoform without HIF-1 alpha activation. PLoS ONE 13(3): e0194782.

Immunoblotting: HIF-2 alpha/EPAS1 Antibody [NB100-122] - CAIX/ HIF-2 alpha/EPAS1 and other antibodies used for immunoblotting to analyze the hypoxia (Hx, 1% O2) or total protein extracts from HeLa cells grown in normoxia (Nx, 21% O2) conditions. Ferecatu I, Canal F, Fabbri L, Mazure NM, Bouton C, Golinelli-Cohen M-P (2018) Dysfunction in the mitochondrial Fe-S assembly machinery leads to formation of the chemoresistant truncated VDAC1 isoform without HIF-1 alpha activation. PLoS ONE 13(3): e0194782.

Reactivity:

Hu, Mu, Rt, Fi, Ha, Pm, Sh

Species Glossary

Reactivity: Hu, Mu, Rt, Fi, Ha, Pm, Sh

Applications:

, , , , , , , , , , , ,

Applications: , , , , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 405	NB100-122AF405
Alexa Fluor 488	NB100-122AF488
Alexa Fluor 647	NB100-122AF647
Alexa Fluor 700	NB100-122AF700
Allophycocyanin/Cy7	NB100-122APCCY7
Biotin	NB100-122B
DyLight 405	NB100-122V
DyLight 488	NB100-122G
DyLight 550	NB100-122R
DyLight 650	NB100-122C
FITC	NB100-122F
HRP	NB100-122H
Janelia Fluor 549	NB100-122JF549
Janelia Fluor 646	NB100-122JF646
PE	NB100-122PE
PE/Atto594	NB100-122PEATT594
PE/Cy5.5	NB100-122PECY55
PE/Cy7	NB100-122PECY7