This protein was produced in an in vitro wheat germ expression system that should preserve correct conformational folding that is necessary for biological function. While it is possible that this protein could display some level of activity, the functionality of this protein has not been explicitly measured or validated.
>80% by SDS-PAGE and Coomassie blue staining
62.5 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.
Read 1 Review rated 4 using H00002597-P02 in the following application:
50 mM Tris-HCI, 10 mM reduced Glutathione, pH 8.0.
>80% by SDS-PAGE and Coomassie blue staining
This product is produced by and distributed for Abnova, a company based in Taiwan.
Alternate Names for Recombinant Human GAPDH Protein
aging-associated gene 9 protein
glyceraldehyde 3-phosphate dehydrogenase
Peptidyl-cysteine S-nitrosylase GAPDH
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
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Peptides and proteins are guaranteed for 3 months from date of receipt.
FAQs for GAPDH Recombinant Protein (H00002597-P02). (Showing 1 - 6 of 6 FAQ).
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.
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.
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.
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.
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.
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.
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.
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.
The GAPDH Antibody: A ChIP Off the Old Block GAPDH antibodies are commonly used as a loading control, due to GAPDH's widespread function as a house-keeping protein. As a leading scientific research antibody supplier, Novus Biologicals offer a wide range of GAPDH antibody reagents and support pro... Read full blog post.
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