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GAPDH Loading Control


GAPDH as Loading Control

GAPDH is commonly used by biological researchers as a loading control for western blot and as a control for qPCR, because the GAPDH gene is often stably and constitutively expressed at high levels in most tissues and cells, which is considered as a housekeeping gene.
However, different regulations of GAPDH under specific conditions have been reported. For example, the transcription factor MZF-1 has been shown to regulate the GAPDH gene. Therefore, the use of GAPDH as loading control has to be considered carefully.

GAPDH Antibody

GAPDH antibody, is usually used as a loading control antibody for Western Blot to normalize the levels of protein detected by confirming that protein loading is the same across the gel. Origene offers the best quality of Anti-GAPDH TrueMAB antibody (Loading control). View all GAPDH loading control antibodies.

GAPDH loading control
Western blot analysis of extracts (35ug) from 9 different cell lines (HepG2: human; HeLa: human; SVT2: mouse; A549: human; COS7: monkey; Jurkat: human; MDCK: canine; PC12: rat; MCF7: human). by using anti-GAPDH monoclonal antibody, Loading Control (TA802519)

GAPDH Molecular Weight

The molecular weight (MW) of GAPDH is approximately 37 kD.

GAPDH Gene

GAPDH gene encodes a member of the glyceraldehyde-3-phosphate dehydrogenase protein family. The encoded protein has been identified as a moonlighting protein based on its ability to perform mechanistically distinct functions. The product of this gene catalyzes an important energy-yielding step in carbohydrate metabolism, the reversible oxidative phosphorylation of glyceraldehyde-3-phosphate in the presence of inorganic phosphate and nicotinamide adenine dinucleotide (NAD). The encoded protein has additionally been identified to have uracil DNA glycosylase activity in the nucleus. Also, this protein contains a peptide that has antimicrobial activity against E. coli, P. aeruginosa, and C. albicans. Studies of a similar protein in mouse have assigned a variety of additional functions including nitrosylation of nuclear proteins, the regulation of mRNA stability, and acting as a transferrin receptor on the cell surface of macrophage. Many pseudogenes similar to this locus are present in the human genome. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Nov 2014]

GAPDH Function

GAPDH has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC. Modulates the organization and assembly of the cytoskeleton. Facilitates the CHP1-dependent microtubule and membrane associations through its ability to stimulate the binding of CHP1 to microtubules (By similarity). Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate. Component of the GAIT (gamma interferon-activated inhibitor of translation) complex which mediates interferon-gamma-induced transcript-selective translation inhibition in inflammation processes. Upon interferon-gamma treatment assembles into the GAIT complex which binds to stem loop-containing GAIT elements in the 3'-UTR of diverse inflammatory mRNAs (such as ceruplasmin) and suppresses their translation. Catalytic Activity: D-glyceraldehyde 3-phosphate + phosphate + NAD+ = 3-phospho-D-glyceroyl phosphate + NADH.

GAPDH References

1. Barber RD, Harmer DW, Coleman RA, Clark BJ (May 2005). "GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues". Physiol. Genomics 21 (3): 389-95.. PMID: 15769908

2. Piszczatowski RT, Rafferty BJ, Rozado A, Tobak S, Lents NH (15 August 2014). "The glyceraldehyde 3-phosphate dehydrogenase gene (GAPDH) is regulated by myeloid zinc finger 1 (MZF-1) and is induced by calcitriol". Biochemical and Biophysical Research Communications 451 (1): 137-141. PMID:25065746

3. "Entrez Gene: GAPDH"

4. "Uniprot: GAPDH"

Loading Control Related Information

Beta-actin loading control GAPDH loading control Beta-tubulin loading control
Loading control FAQs

 

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