Q: Do you offer a smaller size for the GAPDH antibodies?

A: We offer sample sizes for many of our GAPDH loading control antibodies including NB300-221.

Q: I need a suitable loading control antibody that I can use in western blot of rat nerve extracts, which do you recommend?

A: This GAPDH antibody (NB300-221) is a good choice for a loading control antibody and has been cited in over 180 publications and used on rat lysates with good results.

Q: I want to know why GAPDH is used as a loading control antibody in WB techniques.

A: Our GAPDH antibody makes a good loading control because GAPDH is a housekeeping gene essential to metabolism that is globally expressed in most tissue types and cell lines.

Q: I wanted to know which of the two housekeeping genes B-actin or GAPDH can be used for best results. I intend to do an experiment to determine expression of IL-6 and TNF-alpha in liver and kidney tissues.

A: For homogenized tissue, beta-actin and GAPDH are both fine.

Q: I'm looking for a loading control between 24kDa - 65kDa for use in simple western, we tried an actin antibody we already had but it didn't work. What can you recommend?

A: Our GAPDH antibody NB300-221 has been validated in simple western and detects around 35 kDa so I think it would be a good choice for a simple western loading control antibody based on your size requirements.

Q: We need a loading control antibody for WB working with HeLa cell line, will this GAPDH antibody work for that?

A: GAPDH is expressed in HeLa cell lines but whether or not it is the best option for a loading control antibody depends on the size of the protein you are interested in. GAPDH antibody is a good choice for low to mid MW targets, as GAPDH detects around 35 kDa.

Q: We received GAPDH antibody NB600-502 and there was only about 15µl of antibody in the tube. I was under the impression this should have been 100µl.

A: The unit size of this product is 0.1 mg with a concentration of 6.7 mg/ml. Hence the volume should be no less than 14.9 ul.

Q: What secondary antibody should I use for visualization of the GAPDH antibody?

A: This GAPDH antibody is raised in mouse so you would want to use an anti-mouse secondary with the dye of your choice.

GAPDH Antibody (6C5cc) [DyLight 680]

Novus Biologicals | Catalog # NB600-502FR

Submit a review

Citations (13)

Product Documents

Select the "Bulk Orders" button to request additional sizes or formulations.

View Available Conjugates View Available Formulations

100% Guarantee

Product Specifications
Scientific Data
Applications
Preparation & Storage
Background
Product Documents
Specific Notices
Research Areas
Related Blogs

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Porcine, Amphibian, Bovine (Negative), Canine, Chinese Hamster, Feline, Fish, Goat (Negative), Rabbit

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Immunoassay, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation

Cited:

Western Blot

Label

DyLight 680 (Excitation = 692 nm, Emission = 712 nm)

Antibody Source

Monoclonal Mouse IgG₁ Clone # 6C5cc

View all GAPDH Primary Antibodies »

Product Specifications

Immunogen

Hybridoma clone has been derived from hybridization of Sp2/0 myeloma cells with spleen cells of Balb/c mice immunized with Rabbit GAPDH.

Reactivity Notes

Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Additional Mouse on Mouse blocking steps may be required for IHC and ICC experiments. Please contact Technical Support for more information. Chinese Hamster reactivity reported in scientific literature (PMID: 26115091).

Localization

Cytoplasmic, Nucleus, Membrane.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG₁

Theoretical MW

36 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for GAPDH Antibody (6C5cc) [DyLight 680]

GAPDH-Antibody-6C5cc-DyLight-680-Western-Blot-NB600-502FR-img0002.jpg

GAPDH-Antibody-6C5cc-DyLight-680-Western-Blot-NB600-502FR-img0001.jpg

Western Blot: GAPDH Antibody (6C5cc) [DyLight 680] [NB600-502FR] -

Western Blot: GAPDH Antibody (6C5cc) [DyLight 680] [NB600-502FR] - Effective targeting of EcoHIV diminishes post-ischemic stroke inflammation. Mice were infected, treated with ART-7 & ART-11, & subjected to ischemic stroke as in Fig. 6b, c. mRNA levels of cellular activation markers Iba1 (a) & GFAP (b) 7 days post stroke. Protein expression levels of Iba1 (c), GFAP (d), & ICAM1 (e) were quantified by immunoblotting in sham & ischemic stroke animals at day 14 post-stroke in mock (M) & EcoHIV-infected (E) mice that were treated with ART-7 (E + 7) or ART-11 (E + 11). Representative blots are shown, & quantified results are illustrated on the bar graphs. mRNA levels of anti-inflammatory markers ICAM-5 (f) & FoxP3 (g), & tissue degrading enzymes MMP2 (h) & MMP9 (i) were quantified by RT-qPCR. Impact of therapy on viral DNA genome levels was evaluated in ipsilateral hemisphere (j) & spleen (k); n = 5–16 mice per group, 2 independent experiments. Whiskers-box plots represent centerline median, with interquartile range & min-max whiskers. Other graphs represent data as mean & SEM with individual data points. Source data are provided as a Source Data file. *p < 0.05 or **p < 0.01; one-way ANOVA, followed by Tukey multiple comparison test (a, b & f–k), & unpaired t test (c–e) Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31043599), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: GAPDH Antibody (6C5cc) [DyLight 680] [NB600-502FR] -

Western Blot: GAPDH Antibody (6C5cc) [DyLight 680] [NB600-502FR] - Prolonged post-ischemic stroke inflammation in EcoHIV-infected brains. Mice were infected with EcoHIV & subjected to stroke as in Fig. 1. mRNA expression of cytokines IL1 beta (a) & TNF alpha (b) chemokines CXCL1 (c) & CCL2 (d), & cellular activation markers GFAP (e) & Iba1 (f) were assessed by real-time qPCR 7 days post stroke. Sham, n = 6; Stroke, n = 12 mice per group, 3 independent experiments. Time course of GFAP (g) & Iba1 (h) protein expression levels as quantified by western blotting in sham & ischemic stroke animals at days 1, 4, 7, & 14 post-stroke in mock (M) & EcoHIV-infected (E) mice. Representative blots are shown, & quantified results are illustrated on the bar graphs. n = 5–12 mice per group, 3 independent experiments. Whiskers-box plots represent centerline median, with interquartile range & min-max whiskers. Source data are provided as a Source Data file. *p < 0.05; one-way ANOVA, followed by Tukey multiple comparison test (a–f) & unpaired t test (g, h) Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31043599), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: GAPDH Antibody (6C5cc) [DyLight 680] [NB600-502FR] -

Western Blot: GAPDH Antibody (6C5cc) [DyLight 680] [NB600-502FR] - EcoHIV diminishes post-ischemic stroke NVU recovery. Mice were infected with EcoHIV & subjected to stroke as in Fig. 1. Brain sections were stained for laminin (a), ICAM-1 (b) & P-selectin (c) 24 h post stroke, & quantified for mean fluorescence index (MFI); n = 6 mice per group, 10 microvessels per mice, 2 independent experiments. d Time course of ICAM1 expression levels as quantified by western blotting in sham & ischemic stroke animals at days 1, 4, 7, & 14 post-stroke both in mock (M) & EcoHIV-infected (E) mice. Representative blots are shown, & quantified results from 5–12 samples per group are illustrated on the bar graphs. e Representative image (left) & quantified results (right) of infiltration of the infarct area by Lys6g immunoreactive cells (neutrophils) at 24 h post-ischemic stroke. The sections were also stained for MAP2 (neurons) & Hoechst (nuclei). Absence of MAP2 staining indicates infarct area. Data quantified from 6 mice per group, 2 independent experiments, 4 fields of view per mice at ×20 magnification; Z stack images. Whiskers-box plots represent centerline median, with interquartile range & min-max whiskers. Source data are provided as a Source Data file. **p < 0.01; ***p < 0.001; unpaired t test. a–c Scale bars: 40 µm; e scale bar: 320 µm Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31043599), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

View 3 more images