Application DetailsWestern Blotting (WB) and Immunocytochemistry (ICC). A dilution of 1:1,000 is recommended for WB. Human GAPDH has a predicted length of 335 residues and a MW of 36 kDa. A dilution of 1:100 is recommended for IC. Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
SpecificityThe specificity of this antibody has been confirmed by WB. Human, Rat, Mouse, Bovine, Porcine, Chicken
Target Host SpeciesPig
Species ReactivityBovine, Chicken, Human, Mouse, Pig, Rat
Antibody HostMouse
Antibody TypeMonoclonal
Antibody IsotypeIgM
Clone Name1D4
ConjugateUnconjugated
Immunogen DescriptionPurified pig GAPDH
Purity DescriptionPurified
FormatLyophilized from PBS buffer pH 7.2-7.6 with 0.1% trehalose, without preservatives
Reconstitution InstructionsSpin vial briefly before opening. Reconstitute in 250 uL sterile-filtered, ultrapure water. Centrifuge to remove any insoluble material.
Storage InstructionsAfter reconstitution of lyophilized antibody, aliquot and store at -20°C for a higher stability. Avoid freeze-thaw cycles.
Batch NumberPlease see item label.
Expiration Date12 months after date of receipt (unopened vial).
Alternative NamesGlyceraldehyde-3-phosphate dehydrogenase; GAPDH; GAPD; G3PDH; GPDH;
Scientific BackgroundGlyceraldehyde 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. GAPDH may have other roles in the activation of transcription and in the regulation of apoptosis as well as Alzheimer's disease and Huntington's disease. The immunogen used to raise this particular antibody was extensively purified pig GAPDH. 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.
Human neuroblastoma SH-SY5Y cells stained with Mouse monoclonal antibody to Glyceraldehyde 3-Phosphate Dehydrogenase [1D4] M-1376-250 (green), chicken antibody to neurofilament H C-1386-50 (red) and DNA (blue). The antibody reveals strong cytoplasmic staining for GAPDH.
Blots of mice peripheral nerves blotted with M-1376-250 for use as a western blotting control. Note the single sharp band corresponding to GAPDH.
Left: Staining of GAPDH in HeLa cells with mouse anti-GAPDH antibody (1:100, green) by Immunocytochemistry. Blue: DAPI nuclear stain. The GAPDH antibody produces strong cytoplasmic staining of healthy cells. Right: Western blot analysis of cell lysates probed with mouse antibody to GAPDH (1:2,000). [1] protein standard, [2] HEK293, [3] HeLa, [4] SH-SY5Y, [5] COS1, [6] NIH-3T3, and [7] C6. The GAPDH antibody reveals a single band at about 37 kDa in all cell lines. GAPDH is a "house keeping” protein, the level of which is relatively unaffected by most experimental manipulations, and, as a result, this antibody has been widely used as a western blotting loading control.
Specific ReferencesFortun J. et al (2003) Emerging role for autophagy in the removal of aggresomes in Schwann cells. J Neurosci. 2003 Nov 19;23(33):10672-80. Felitsyn N. et al (2008) The heme precursor delta-aminolevulinate blocks peripheral myelin formation. J Neurochem. 2008 Sep;106(5):2068-79. Iskandar M. et al (2005) Copper chaperone for Cu/Zn superoxide dismutase is a sensitive biomarker of mild copper deficiency induced by moderately high intakes of zinc. Nutr J. 2005 Nov 24;4:35. Bizzozero O.A. et al (2009) Identification of major S-nitrosylated proteins in murine experimental autoimmune encephalomyelitis. J Neurosci Res. 2009 Oct;87(13):2881-9. Madorsky I. et al (2009) Intermittent fasting alleviates the neuropathic phenotype in a mouse model of Charcot-Marie-Tooth disease. Neurobiol Dis. 2009 Apr;34(1):146-54. Bertinato J. et al (2010) Decreased Erythrocyte CCS Content is a Biomarker of Copper Overload in Rats. Int J Mol Sci. 2010 Jul 2;11(7):2624-35. Rangaraju S. et al (2008) Pharmacological induction of the heat shock response improves myelination in a neuropathic model. Neurobiol Dis. 2008 Oct;32(1):105-15. Rangaraju S. et al (2009) Molecular architecture of myelinated peripheral nerves is supported by calorie restriction with aging. Aging Cell. 2009 Apr;8(2):178-91.
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