BACKGROUND GAPDH (Glyceraldehyde 3-phosphate dehydrogenase, G3PDH) is an enzyme that catalyzes the reversible Oxidative phosphorylation of glyceraldehyde-3-phosphate in the presence of inorganic phosphate and nicotinamide adenine dinucleotide (NAD), the sixth step of glycolysis and thus serves to break down glucose for energy and carbon molecules.1 In addition to this long established metabolic function, GAPDH has recently been implicated in several non-metabolic processes, including transcription activation, initiation of apoptosis, and ER to Golgi vesicle shuttling.2 The enzyme exists as a tetramer of identical chains.
Because the GAPDH gene is often stably and constitutively expressed at high levels in most tissues and cells, it is considered a housekeeping gene. For this reason, GAPDH is commonly used by biological researchers as a loading control for western blot and as a control for RT-PCR. However, many researchers report different regulation of GAPDH under specific conditions.3 Therefore, the use of GAPDH as loading control has to be controlled carefully.
REFERENCES
1. Voet, D. & Voet, J. G.: Biochemistry, Third Edition. J. Wiley & Sons, Hoboken, NJ, 2004
2. Tarze, A. et al: Oncogene 26:2606-20, 2007
3. Graven, K.K. et al: J. Biol. Chem. 269:24446-53, 1994
Products are for research use only. They are not intended for human, animal, or diagnostic applications.
Cat.No.: | CP10112 |
Antigen: | Raised against purified recombinant human GAPDH fragments expressed in E. coli. |
Isotype: | Mouse IgG1 |
Species & predicted species cross- reactivity ( ): | Human, Mouse, Rat |
Applications & Suggested starting dilutions:* | WB 1:1000 ELISA 1:10000 IHC 1:200 ICC 1:200 |
Predicted Molecular Weight of protein: | 35 kDa |
Specificity/Sensitivity: | Detects endogenous GAPDH proteins without cross-reactivity with other related proteins |
Storage: | Store at -20°C, 4°C for frequent use. Avoid repeated freeze-thaw cycles. |
*Optimal working dilutions must be determined by end user.