GAPDH Antibody

Product Discontinued

GAPDH Antibody Summary

• Immunogen: Amino acids 73-99 of glyceraldehyde 3-phosphate dehydrogenase protein were used as the immunogen for this antibody.
• Marker: Cytosolic Marker
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: GAPDH
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Western Blot

Reactivity Notes

The amino acid sequence used as immunogen is 100% homologous in human, cattle, rat, mouse, chimpanzee, rhesus monkey, opossum, pig, chicken and dog, and 94% homologous in goldfish.

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: GAPDH antibody at 0.5 mg/ml in PBS containing 0.05% BSA
• Preservative: 0.05% Sodium Azide
• Concentration: 0.5 mg/ml
• Purity: Protein G purified

Alternate Names for GAPDH Antibody

• aging-associated gene 9 protein
• EC 1.2.1
• EC 1.2.1.12
• EC 2.6.99.-
• G3PD
• G3PDH
• GAPD
• GAPDH
• glyceraldehyde 3-phosphate dehydrogenase
• glyceraldehyde-3-phosphate dehydrogenase
• MGC88685
• Peptidyl-cysteine S-nitrosylase GAPDH

Background

Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) is a ubiquitous enzyme involved in glycolysis, converting glyceraldehyde-3-phosphate into 1,3 diphosphoglycerate. This constitutively expressed, homotetramer protein can be found in the nucleus and cytoplasm, and the monomer has a theoretical molecular weight of 36 kDa. Known as a housekeeping gene, GAPDH routinely serves as a control for real-time PCR (RT-PCR) and as a loading control for Western Blots (1-3). In addition to its role in glycolysis, GAPDH has multiple cellular functions including membrane trafficking, transcription activation, apoptosis, DNA repair, and has been implicated in various pathologies such as cancer and neurodegenerative diseases including Alzheimer's and Huntington's (4,5).

References

1) Barber RD, Harmer DW, Coleman RA, Clark BJ. (2005) GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues. Physiol Genomics. 21(3):389-95. PMID: 15769908

2) Jia Y, Takimoto K. (2006) Mitogen-activated protein kinases control cardiac KChIP2 gene expression. Circ Res. 98(3):386-93. PMID: 16385079

3) Godsel LM, Hsieh SN, Amargo EV, Bass AE, Pascoe-McGillicuddy LT, Huen AC, Thorne ME, Gaudry CA, Park JK, Myung K, Goldman RD, Chew TL, Green KJ. (2005) Desmoplakin assembly dynamics in four dimensions: multiple phases differentially regulated by intermediate filaments and actin. J Cell Biol. 171(6):1045-59. PMID: 16365169

4) Sirover MA1. (1999) New insights into an old protein: the functional diversity of mammalian glyceraldehyde-3-phosphate dehydrogenase. Biochim Biophys Acta. 1432(2): 159-84. PMID: 10407139

5) Tristan C, Shahani N, Sedlak TW, Sawa A. (2011) The diverse functions of GAPDH: views from different subcellular compartments. Cell Signal. 23(2):317-23. PMID: 20727968

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.

Publications for GAPDH Antibody (NBP2-24396)(15)

Publications using NBP2-24396

• Shi M, Han W, Spivack SD. A quantitative method to identify microRNAs targeting a messenger RNA using a 3'UTR RNA affinity technique. Anal Biochem. 2013-08-09 [PMID: 23938772]
• Du H, Huang Y, Zaghlula M et al. The MID1 E3 ligase catalyzes the polyubiquitination of Alpha4 (a4), a regulatory subunit of protein phosphatase 2A (PP2A): novel insights into MID1-mediated regulation of PP2A. J Biol Chem. 2013-07-19 [PMID: 23740247]
• Chauhan SK, Singh VV, Rai R et al. Distinct autoantibody profiles in systemic lupus erythematosus patients are selectively associated with TLR7 and TLR9 upregulation. J Clin Immunol. 2013-07-01 [PMID: 23564191]
  
  Details:
  PBLs (WB) or B-cells (Flow intracellular) from SLE patients or controls: 1. TLR7 mAb clone IMG4G6 (IMG-6675A): WB (Figs 2a, 2c).
• Saha T. LAMP2A overexpression in breast tumors promotes cancer cell survival via chaperone-mediated autophagy. Autophagy. 2012-11-01 [PMID: 22874552]
• Roy S, Short MK, Stanley ER, Jubinsky PT. Essential role of Drosophila black-pearl is mediated by its effects on mitochondrial respiration. FASEB J. 2012-09-01 [PMID: 22700875]
• Mikami T, Hada T, Chosa N et al. Expression of Wilms' tumor 1 (WT1) in oral squamous cell carcinoma. J Oral Pathol Med. 2013-02-01 [PMID: 22672247]
• Hart AK, Fioravante D, Liu RY et al. Serotonin-mediated synapsin expression is necessary for long-term facilitation of the Aplysia sensorimotor synapse. J Neurosci. 2011-12-14 [PMID: 22171042]
• Lai WT, Krishnappa V, Phinney DG. Fibroblast growth factor 2 (Fgf2) inhibits differentiation of mesenchymal stem cells by inducing Twist2 and Spry4, blocking extracellular regulated kinase activation, and altering Fgf receptor expression levels. Stem Cells. 2011-07-01 [PMID: 21608080]
• Rajaraman G, Murthi P, Brennecke SP, Kalionis B. Homeobox gene HLX is a regulator of HGF/c-met-mediated migration of human trophoblast-derived cell lines. Biol Reprod. 2010-10-01 [PMID: 20554918]
  
  Details:
  Product cited: Western Blot Loading Control Kit (IMG-6166K); GAPDH (IMG-5143, component) was used in WB in Human SGHPL-4 (trophoblast) and HTR-8/SVneo cells, Fig 7. Matrix metalloproteinases -8, -9, and -12 in smokers and patients with Stage 0 COPD. Illum
• Tan XL, Shi M, Tang H et al. Candidate dietary phytochemicals modulate expression of phase II enzymes GSTP1 and NQO1 in human lung cells. J Nutr. 2010-08-01 [PMID: 20554899]
• Miller CL, Dulay JR. The high-affinity niacin receptor HM74A is decreased in the anterior cingulate cortex of individuals with schizophrenia. Brain Res Bull. 2008-09-05 [PMID: 18639743]
• Wagner MW, Li LS, Morales JC et al. Role of c-Abl kinase in DNA mismatch repair-dependent G2 cell cycle checkpoint arrest responses. J Biol Chem. 2008-08-01 [PMID: 18480061]
• Xiong X, Cui P, Hossain S et al. Allosteric inhibition of the nonMyristoylated c-Abl tyrosine kinase by phosphopeptides derived from Abi1/Hssh3bp1. Biochim Biophys Acta. 2008-05-01 [PMID: 18328268]
• Takagi S, Nakajima M, Mohri T, Yokoi T. Post-transcriptional regulation of human pregnane X receptor by micro-RNA affects the expression of cytochrome P450 3A4. J Biol Chem. 2008-04-11 [PMID: 18268015]
• Baskar S, Kwong KY, Hofer T et al. Unique cell surface expression of receptor tyrosine kinase ROR1 in human B-cell chronic lymphocytic leukemia. Clin Cancer Res. 2008-01-15 [PMID: 18223214]

FAQs for GAPDH Antibody (NBP2-24396). (Showing 1 - 7 of 7 FAQs).

1. We need a loading control antibody for WB working with HeLa cell line, will this GAPDH antibody work for that?
   • 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.
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.
7. 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.
   • For homogenized tissue, beta-actin and GAPDH are both fine.

Bioinformatics

• Gene Symbol: GAPDH