The GAPDH (glyceraldehyde-3-phosphate dehydrogenase) gene is mapped to human chromosome 12p13.3 and encodes a tetramer containing identical chains. The enzyme is commonly known as glycolytic enzyme and contains a binding site for NAD+(nicotinamide adenine diphosphate) and glyceraldehyde-3-phosphate. GAPDH contributes to 10-20% of the total cellular protein and is considered to be evolutionarily conserved.
Anti-GAPDH recognizes human, mouse, and rat GAPDH.
Synthetic peptide corresponding to amino acids of mouse GAPDH, conjugated to KLH via an N-terminal cysteine residue. The corresponding sequence is identical in rat and differs by two amino acids in humans.
Anti-GAPDH antibody produced in rabbit is suitable as a primary antibody for western blotting using:
It is suitable for immunoprecipitation using mouse NIH3T3 cell lysates with 5-10μg, for indirect immunofluorescence using rat NRK cells at a working concentration of 5-10μg/mL. It is also suitable for western blotting at a working concentration of 0.1-0.2μg/mL using a whole extract of human HeLa cells.
- human cancer cell extract
- extract from muscle tissue from old mice exhibiting sarcopenia
- trabeculae or samples of the mice heart myocardium
- mouse embryonic fibroblasts
The enzyme glyceraldehyde-3-phosphate dehydrogenase catalyzes the reversible oxidative phosphorylation of glyceraldehyde-phosphate in the presence of inorganic phosphate and NAD, which is a critical energy-yielding step in carbohydrate metabolism. It binds to several proteins including actin, tubulin, amyloid precursor, polyglutamine peptides, DRPLA (dentatorubral-pallidoluysian atrophy), and huntingtin. GAPDH forms a part of OCA-S, the multicomponent OCT1 (octamer-motif-binding factor) coactivator complex, which is involved in the S phase-dependent histone H2B transcription. This association is responsible for linking H2B transcriptional machinery to cell cycle regulation and to the cellular metabolic state. In response to oxidative stress, GAPDH induces apoptosis.
Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide.
Storage and Stability
For continuous use, store at 2-8 °C for up to one month.
For extended storage, freeze in working aliquots. Repeated freezing and thawing, or storage in “frost free” freezers, is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilutions should be discarded if not used within 12 hours.
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