Catalogue# MCA-1D4: Mouse Monoclonal Antibody to Glyceraldehyde
3-Phosphate Dehydrogenase (GAPDH, G3PDH or GPDH)
The Immunogen: Glyceraldehyde 3-Phosphate Dehydrogenase
(GAPDH) is a metabolic enzyme responsible for catalyzing one step in the
glycolytic pathway, the reversible oxidative phosphorylation of glyceraldehyde
3-phosphate. Because GAPDH as a protein expressed in large amounts and which
is required at all times for an important "house keeping" functions,
levels of GAPDH mRNA are often measured and used as standards in studies
of mRNA expression. Increasingly, scientists are making use of specific
antibodies to GAPDH in comparable studies of levels of protein expression.
This antibody can be used as a loading control for western blotting experiments,
allowing comparison between the level of this protein and others in a cell
or tissue. Apart from a role in glycolysis, GAPDH may have other roles such
as in the activation of transcription (1). GAPDH is reported to bind to
a variety of other proteins, including the amyloid precursor protein, mutations
in which cause some forms of Alzheimer's disease, and the polyglutamine
tracts of Huntingtin, the protein product aberrant forms of which are causative
of Huntington's disease (2,3). Associations with actin and tubulin have
also been reported. The protein may also have a role in the regulation of
apoptosis, and interestingly migrates from the cytoplasm into the nucleus
when cells become apoptotic (4). The immunogen used to raise this particular
antibody was extensively purified pig GAPDH.
Left: blots of mice peripheral nerves blotted with MCA-1D4 for use as an western blotting control. Note the single sharp band corresponding to GAPDH. Right: Human neuroblastoma SH-SY5Y cells stained with MCA-1D4 (green), chicken antibody to neurofilament NF-H CPCA-NF-H (red) and DNA (blue). The antibody reveals strong cytoplasmic staining for GAPDH.
Antibody characteristics: MCA-1D4 is a mouse IgM class antibody with a k light chain. IgM antibodies differ from IgGs in that they contain a total of ten antigen binding sites instead of only two. They can be used in the same was as IgGs, and most secondary reagents binding IgGs will also bind IgMs. MCA-1D4 recognizes GAPDH specifically both in western blots and in immunocytochemical experiments. On blots MCA-1D4 reveals a prominent ~38kDa band, and on cells in tissue culture the antibody stains in a punctate cytoplasmic fashion, for images press here. MCA-1D4 is known to react with GAPDH from human, cow, pig, mouse, rat and other mammals, and also recognizes avian GAPDH. Since GAPDH is one of the most conserved proteins known, it is likely that the antibody is effective on other species also.
Suggestions for use: The antibody solution was made using Integra CL-350 cells and is therefore concentrated tissue culture supernatant, with an antibody concentration of at least 1mg/ml. The antibody solution can be used at dilutions of at least 1:100 in immunofluorescence experiments. In western blotting using chemiluminescence it can be used at dilutions of 1:1,000 or lower. It is very useful as a loading control for quantitative western blotting (see for example references 5 and 6). Antibody preparation contains 10mM sodium azide preservative (Link to http://www.encorbio.com/MSDS/azide.htm for Material Safety Data Sheet). Avoid repeated freezing and thawing, store at 4°C or -20°C.
Limitations: This product is for research use only and is not approved for use in humans or in clinical diagnosis.
Omim link: press here
References:
1. Morgenegg G, Winkler GC, Hubscher U, Heizmann CW, Mous J, Kuenzle CC. Glyceraldehyde-3-phosphate dehydrogenase is a nonhistone protein and a possible activator of transcription in neurons. J Neurochem. 47:54-62 1986
2. Schulze H, Schuler A, Stuber D, Dobeli H, Langern H & Huber G. Rat brain glyceraldehyde-3-phosphate dehydrogenase interacts with the recombinant cytoplasmic domain of Alzheimer's beta-amyloid precursor protein. J Neurochem. 60:1915-22 1993
3. Burke
JR, Enghild JJ, Martin ME, Jou Y-S, Myers RM, Roses AD, Vance JM & Strittmatter
WJ. Huntingtin and DRPLA proteins selectively interact with the enzyme GAPDH. Nature
Med. 2: 347-350, 1996.
4. Dastoor Z. & Dreyer, J-L. Potential role of nuclear translocation of
glyceraldehyde-3-phosphate dehydrogenase in apoptosis and oxidative stress. J.
Cell Sci. 114:1643-1653 2001.
5. Fortun J, Dunn WA, Joy S, Li J. & Notterpek, L. Emerging Role for Autophagy in the Removal of Aggresomes in Schwann Cells. J. Neurosci. 23:10672-10680 2003.
6. Ellis RC, Earnhardt JN, Hayes RL, Wang KK & Anderson DK. Cathepsin B mRNA and protein expression following contusion spinal cord injury in rats. J Neurochem. 88:689-97 2004.
7. Fortun J, Li J, Go J, Fenstermaker A, Fletcher BS & Notterpek L. Impaired proteasome activity and accumulation of ubiquitinated substrates in a hereditary neuropathy model. J Neurochem. 92:1531-41 2005
8. Fortun J, Li J, Go J, Fenstermaker A, Fletcher BS & Notterpek L. Impaired proteasome activity and accumulation of ubiquitinated substrates in a hereditary neuropathy model. J Neurochem. 93:766-8 2005
9. Iskandar M, Swist E, Trick KD, Wang B, L'Abbe MR, Bertinato J. Copper chaperone for Cu/Zn superoxide dismutase is a sensitive biomarker of mild copper deficiency induced by moderately high intakes of zinc. Nutr J. 4:35 2005
10. Fortun J, Go JC, Li J, Amici SA, Dunn WA Jr, Notterpek L. Alterations in degradative pathways and protein aggregation in a neuropathy model based on PMP22 overexpression. Neurobiol Dis. 22:153-164 2006
11. Amici SA, Dunn WA, Murphy AJ, Adams NC, Gale NW, Valenzuela DM, Yancopoulos GD & Notterpek L.Peripheral Myelin Protein 22 Is in Complex with {alpha}6beta4 Integrin, and Its Absence Alters the Schwann Cell Basal Lamina. J. Neurosci. 26:1179-89 2006
12. Amici SA, Dunn WA & Notterpek, L. Developmental abnormalities in the nerves of peripheral myelin protein 22-deficient mice. J. Neurosci. Res. 85:238-249 2006
13. Felitsyn N, Stacpoole, PW & Nottepek L. Dichloroacetate causes reversible demyelination in vitro: potential mechanism for its neuropathic effect. J. Neurochem 100:429-36 2007
Examples: For some on line images generated with this antibody press here
OMIM link: Press here
Availability and Price: Available for shipping now, $200 US per aliquot of 0.5mls of concentrated tissue culture supernatant, enough for hundreds of experiments. For order form press here