GAPDH Antibody (6C5)

Images

 
Western Blot: GAPDH Antibody (6C5) [NB600-502] - Western blot analysis of GAPDH expression in 1) HeLa, 2) NTERA-2, 3) A-431, 4) HepG2, 5) MCF-7, 6) NIH 3T3, 7) PC-12 and 8) COS-7 whole cell lysates.
Simple Western: GAPDH Antibody (6C5) [NB600-502] - Simple Western lane view shows a specific band for GAPDH in 0.5 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Po, Am, Ca, Fe, Fi, Mk, Pm, Rb, Bv(-), Gt(-)Species Glossary
Applications WB, Simple Western, IA, ICC/IF
Clone
6C5
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated

GAPDH Antibody (6C5) Summary

Immunogen
Hybridoma clone has been derived from hybridization of Sp2/0 myeloma cells with spleen cells of Balb/c mice immunized with human or rabbit GAPDH.
Localization
Cytoplasmic, Nucleus, Membrane.
Isotype
IgG1
Clonality
Monoclonal
Host
Mouse
Gene
GAPDH
Purity
Protein A purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 0.5-1 ug/ml
  • Simple Western 1:100
  • Immunoassay
  • Immunocytochemistry/Immunofluorescence 1:10-1:2000
Application Notes
Since GAPDH is expressed in all cells it is for example becoming the marker of choice for a loading control in Western Blotting.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
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.
Positive Control
A-431 Lysate (NB800-PC12)
HeLa Lysate (NB800-PC1)
MCF-7 Lysate (NB820-59465)
GAPDH Lysate (NBL1-10967)
HepG2 Lysate (NBP1-42569)
NTERA-2 Lysate (NBP1-44968)
PC-12 Lysate (NBP2-13170)
Reviewed Applications
Read 1 Review rated 5
using
NB600-502 in the following applications:

Publications
Read Publications using
NB600-502 in the following applications:

  • 1 publication
  • WB
    22 publications

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.

Packaging, Storage & Formulations

Storage
Store at 4C. Do not freeze.
Buffer
PBS (pH 7.4)
Preservative
0.09% Sodium Azide
Purity
Protein A purified

Alternate Names for GAPDH Antibody (6C5)

  • aging-associated gene 9 protein
  • anti-GAPDH 6c5
  • EC 1.2.1
  • EC 1.2.1.12
  • EC 2.6.99.-
  • G3PD
  • G3PDH
  • gapdh 6c5
  • GAPDH
  • GAPDPeptidyl-cysteine S-nitrosylase GAPDH
  • glyceraldehyde 3-phosphate dehydrogenase
  • glyceraldehyde-3-phosphate dehydrogenase
  • MGC88685

Background

Glyceraldehyde 3 phosphate dehydrogenase (GAPDH) is well known as one of the key enzymes involved in glycolysis. Besides its functioning as a glycolytic enzyme in cytoplasm, recent evidence suggests that mammalian GAPDH is also involved in a great number of intracellular proceses such as membrane fusion, microtubule bundling, phosphotransferase activity, nuclear RNA export, DNA replication, and DNA repair. Recently data has implicated GAPDH in different pathologies including prostate cancer progression, programmed neuronal cell death, age-related neuronal diseases, such as Alzheimer's and Huntington's disease. Some physiological factors, such as hypoxia and diabetes, increase GAPDH expression in certain cell types.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Publications for GAPDH Antibody (NB600-502)(27)

We have publications tested in 5 confirmed species: Human, Mouse, Rat, Amphibian, Chinese hamster.

We have publications tested in 2 applications: ICC/IF, WB.


Filter By Application
ICC/IF
(1)
WB
(22)
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Filter By Species
Human
(12)
Mouse
(7)
Rat
(1)
Amphibian
(1)
Chinese hamster
(1)
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Showing Publications 1 - 10 of 27. Show All 27 Publications.
Publications using NB600-502 Applications Species
Tacconi EM, Lai X, Folio C et al. BRCA1 and BRCA2 tumor suppressors protect against endogenous acetaldehyde toxicity EMBO Mol Med 2017 Jul 20 [PMID: 28729482] (WB, Mouse) WB Mouse
Lai X, Broderick R, Bergoglio V et al. MUS81 nuclease activity is essential for replication stress tolerance and chromosome segregation in BRCA2-deficient cells Nat Commun 2017 Jul 17 [PMID: 28714477] (WB, Human) WB Human
Jing R, Duncan CB, Duncan SA. A small molecule screen reveals that HSP90B promotes the conversion of iPSC-derived endoderm to a hepatic fate and regulates HNF4A turnover. Development. Mar 30 2017 [PMID: 28360131]
Shao Z, Lee X, Huang G et al. LINGO-1 Regulates Oligodendrocyte Differentiation through the Cytoplasmic Gelsolin Signaling Pathway. J. Neurosci. 2017 Feb 13 [PMID: 28193690] (WB, Human) WB Human
Zhang P, Tsuchiya K, Kinoshita T et al. Vitamin B6 prevents IL-1B production by inhibiting NLRP3 inflammasome activation. J. Biol. Chem. Oct 12 2016 [PMID: 27733681] (WB, Mouse) WB Mouse
Giorgetti E, Yu Z, Chua Jp et al. Rescue of Metabolic Alterations in AR113Q Skeletal Muscle by Peripheral Androgen Receptor Gene Silencing. Cell Rep Sep 27 2016 [PMID: 27681426]
Baik Ih, Jo Gh, Seo D et al. Knockdown of RPL9 expression inhibits colorectal carcinoma growth via the inactivation of Id-1/NF-kB signaling axis. Int. J. Oncol. Sep 13 2016 [PMID: 27633352] (WB) WB
Ashton TM, Fokas E, Kunz-Schughart LA et al. The anti-malarial atovaquone increases radiosensitivity by alleviating tumour hypoxia Nat Commun 2016 Jul 25 [PMID: 27453292] (WB) WB
Cervantes-Gomez F, Kumar Patel V, Bose P et al. Decrease in total protein level of Bruton's tyrosine kinase during ibrutinib therapy in chronic lymphocytic leukemia lymphocytes. Leukemia 2016 Aug [PMID: 27198052]
Tavori H, Giunzioni I, Predazzi IM et al. Human PCSK9 promotes hepatic lipogenesis and atherosclerosis development via apoE- and LDLR-mediated mechanisms. Cardiovasc. Res. May 15 2016 [PMID: 26980204] (WB, Mouse) WB Mouse
Show All 27 Publications.

Review for GAPDH Antibody (NB600-502) (1) 51

Average Rating: 5
(Based on 1 review)
We have 1 review tested in 1 species: Human.
We have 1 review tested in 1 application: WB.

Reviews using NB600-502:
Filter by Applications
WB
(1)
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Filter by Species
Human
(1)
All Species
Images Ratings Applications Species Date Details
Western Blot GAPDH NB600-502
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reviewed by:
Anonymous
WB Human 06/17/2014
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Summary

ApplicationWestern Blot
Sample Tested293T and HepG2 lysates
SpeciesHuman

Blocking

Blocking Details5% BSA in PBS

Primary Anitbody

Dilution Ratio1:2000 diluted in PBS with triton X-100, blot was incubated at room temperature on a rocker

Secondary Antibody

Secondary Descriptionanti-mouse IgG HRP conjugate (Jackson Labs)
Secondary Manufacturer Cat#115-005-174
Secondary Concentration1:15000

Details

Detection NotesProbed using Visualizer Western Blot Detection Kit (Millipore) and captured on X-ray films, 5 sec exposure

Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for GAPDH Antibody (NB600-502). (Showing 1 - 6 of 6 FAQs).

  1. We need a loading control antibody for WB working with HeLa cell line, will this GAPDH antibody work for that?
    • Well 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 lookin at. GAPDH antibody is a good choice for low to mid MW targets as GAPDH detects around 35 kDa.
  2. I want to know why GAPDH is used as a loading control antibody in WB techniques.
    • 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.
  3. I need a suitable loading control antibody that I can use in western blot of rat nerve extracts, which do you recommend?
    • 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.
  4. 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?
    • 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.
  5. What secondary antibody should I use for visualization of the GAPDH antibody?
    • This GAPDH antibody is raised in mouse so you would want to use an anti-mouse secondary with the dye of your choice.
  6. Do you offer a smaller size for the GAPDH antibodies?
    • We offer sample sizes for many of our GAPDH loading control antibodies including NB300-221.

Positive Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NB600-502AF405
Alexa Fluor 488 Labeled NB600-502AF488
Alexa Fluor 647 Labeled NB600-502AF647
Alexa Fluor 700 Labeled NB600-502AF700
DyLight 350 Labeled NB600-502UV
DyLight 405 Labeled NB600-502V
DyLight 488 Labeled NB600-502G
DyLight 680 Labeled NB600-502FR
DyLight 755 Labeled NB600-502IR

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Bioinformatics Tool for GAPDH Antibody (NB600-502)

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Blogs on GAPDH. Showing 1-10 of 12 blog posts - Show all blog posts.

The use of Beta Actin (AC-15) as a loading control across multiple species
Actin is a fundamental component of the cytoskeleton, where it has the ability to create and break down actin filament formation in response to various cell needs.  Actin has six highly conserved isoforms, however beta and gamma actin are the two...  Read full blog post.

Tips on choosing an ideal loading control antibody for Western Blotting
Western blotting is one of the most commonly used antibody assay techniques in cell and molecular biology research since its development over three decades ago, and is considered the gold standard for protein detection and quantification. When p...  Read full blog post.

GAPDH - A "housekeeping" gene with diverse functions in cellular homeostasis
Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is a well-known housekeeping gene with functions in glycolysis. Many biologists are familiar with the gene and use GAPDH antibodies for a loading control when performing west blots. However, this pri...  Read full blog post.

A New Standard in Antibody Testing - Simple Western Certified Antibodies
The Western blot is one of the most commonly used antibody assay techniques in cell and molecular biology research since its development over three decades ago, and is considered the gold standard for protein detection and quantification. The tradi...  Read full blog post.

GAPDH: More than a housekeeping gene
GAPDH is a 146kD tetramer glycolytic pathway metabolic enzyme composed of four 30-40 kDa subunits. It is responsible for reversibly phosphorylating its substrate glyceraldehyde 3-phosphate within the glycolytic pathway.  Apart from its role in gl...  Read full blog post.

GAPDH (Glyceraldehyde 3-Phosphate Dehydrogenase)
GAPDH is a 146 kD tetramer glycolytic pathway metabolic enzyme responsible for reversibly phosphorylating glyceraldehyde 3-phosphate. It may have other possible functions in transcriptional activation. GAPDH is highly expressed due to this housekeepin...  Read full blog post.

Time to Start Actin Like a Reliable 'Housekeeper'!
A growing body of data and studies using actin antibodies supports a view of the actin cytoskeleton of smooth muscle cells as a dynamic structure that plays an integral role in regulating the development of mechanical tension and the material properti...  Read full blog post.

Beta Actin and GAPDH: The Importance of Loading Controls
Western blotting is an essential technique to probe protein expression in complex cell or tissue lysates. To accurately determine protein expression and interpret Western blot results, it is important to use loading controls. A loading control anti...  Read full blog post.

The GAPDH Antibody in Western blot Assays
The loading controls on our antibody database are widely used in gel electrophoresis and Western blotting studies. Products like the GAPDH antibody detect "housekeeping" proteins which are abundantly distributed in cells. This makes them useful for ch...  Read full blog post.

New Primers Available for ChIP Procedures
Novus has recently released six new primer sets designed for use in real-time PCR DNA amplification of housekeeping, silent, or heterochromatin associated proteins. Specifically, the primers are for GAPDH, RPL30, MyoD1, AFM, alpha Satellite, and Satel...  Read full blog post.

Showing 1-10 of 12 blog posts - Show all blog posts.

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Recent Reviews

5
5
1
4
0
3
0
2
0
1
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Anonymous
06/17/2014
Application: WB
Species: Human

Bioinformatics

Gene Symbol GAPDH
Entrez
Uniprot