Application DetailsWestern Blotting (WB) and Immunocytochemistry (ICC). A dilution of 1:10,000-1:30,000 is recommended for WB. A dilution of 1:1,000-1:10,000 is recommended for ICC. Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
SpecificitySpecific for the ~36-38 kDa Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) protein.
Target Host SpeciesHuman
Species ReactivityHuman, Mouse, Other Mammals (Predicted)
Antibody HostRabbit
Antibody TypePolyclonal
Antibody IsotypeIgG
ConjugateUnconjugated
Immunogen DescriptionFull length recombinant human GAPDH expressed in and purified from E. coli
Purity DescriptionWhole serum
FormatLyophilized, without preservatives.
Reconstitution InstructionsSpin vial briefly before opening. Reconstitute in 100 µL 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.
HeLa cell extract blotted with rabbit anti-GAPDH. Note the single sharp band corresponding to GAPDH.
HeLa cells stained with rabbit anti-GAPDH (red), monoclonal to TAF15 (green) and DNA (blue). The antibody reveals strong cytoplasmic staining for GAPDH.
Left: HeLa cells stained with rabbit antibody to GAPDH (red, 1:2,000) by Immunocytochemistry. Blue: Hoechst nuclear stain. The GAPDH antibody produces diffuse cytoplasmic staining. Right: Western blot analysis of cell cytosolic- or nuclear-enriched fractions, using rabbit antibody to GAPDH (red, 1:20,000, lanes 2-5). [1] protein standard, [2] NIH-3T3 cytosolic, [3] NIH-3T3 nuclear, [4] HeLa cytosolic, and [5] HeLa nuclear fractions. A strong band at 37 kDa corresponds to GAPDH protein, mainly detected in the cytosolic fractions. The same blot was simultaneously probed with mouse antibody to the nuclear RNA binding protein SF3B4 (M-1576-100, green, 1:1,000). In contrast to GAPDH, SF3B4 is exclusively expressed in the nuclear fraction.
General ReferencesMorgenegg 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 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 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. 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 200100:429-36 (2007).
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