Background[1-3]
Glyceraldehyde 3-phosphate dehydrogenase monoclonal antibody (internal reference antibody) is an antibody protein that can specifically bind to glyceraldehyde 3-phosphate dehydrogenase. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is a key enzyme in the glycolysis process. In addition to serving as a glycolytic enzyme in the cytoplasm, there is evidence that GAPDH in mammalian cells participates in a variety of intracellular biochemical processes, including membrane fusion, microtubule bundling, and phosphotransfer Enzyme (phosphotransferase) activation, RNA export from the nucleus, DNA replication and DNA repair. Some physiological factors, such as hypoxia and diabetes, can increase the expression of GAPDH in specific cells. GAPDH is present in almost all tissues and is continuously expressed at high levels.
GAPDH (glyceraldehyde-3-phosphate dehydrogenase) is a key enzyme involved in glycolysis and consists of four 30-40kDa subunits. xy-13254R GAAα-glucosidase/lysosomal α-glucosidase antibody GAPDH protein is expressed at high levels in almost all tissues and is widely used as an internal reference for Western blot protein standardization. It is a good internal reference antibody. As a housekeeping gene, the protein expression level of GAPDH is generally constant in the same cells or tissues. In the experiment, there may be inaccurate determination of total protein concentration; or operational errors between samples caused when protein samples are loaded before electrophoresis; these errors need to be determined by measuring the content of GAPDH actually transferred to the membrane in each sample. Calibration, so general western experiments require internal reference settings. The specific calibration method is to divide the measured target protein content of each sample by the GAPDH content of this sample to obtain the relative content of the target protein of each sample. Only then are sample-to-sample comparisons made.
Apply[4][5]
For research on GAPDH as a universal internal standard protein in prokaryotes and eukaryotes and related biotechnology research and development
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a very conserved and important multifunctional enzyme in organisms, which is widely found in prokaryotes and eukaryotes. , such as Escherichia coli, yeast, Caenorhabditis elegans, rats and mice and other model organisms, mainly participate in the sixth step of glycolysis in the body’s sugar metabolism, that is, catalyze the conversion of glyceraldehyde-3-phosphate into 1,3-di The process of phosphoglycerate and releasing energy (ATP). The molecular weight of GAPDH monomers in the biological world is between 30kD and 40kD. GAPDH in its natural state mainly exists in the form of tetramers. Its amino acid coding sequence is highly conserved in both prokaryotes and eukaryotes, with a homology of up to 76%. The GAPDH protein in prokaryotes and eukaryotes is an important protease superfamily with very conserved functions. It is encoded by different genes in different species, such as the gapA, gapB and gapC genes of microorganisms, TDH-1 of yeast, TDH-2 and TDH-3 genes, gpd-1, gpd-2, gpd-3 and gpd-4 genes of Caenorhabditis elegans, G3PDH gene of rats, etc. The existence of a variety of coding genes in different species on the one hand suggests the functional compensatory nature of different GAPDH variants, and on the other hand reflects the high degree of regulation of individuals in response to environmental stress, providing evidence for their functional conservation as housekeeping proteins. important basis.
Internal standard proteins are an important type of housekeeping proteins in organisms, with high sequence conservation and wide tissue distribution. Common internal standard proteins include: cytoskeletal protein actin, tubulin Tubulin, cell metabolism-related protein GAPDH, etc. Among them, the protein expression level of GAPDH in the same type of cells or tissues is relatively constant and is rarely affected by external stress factors. It has been widely used as an internal reference molecule for the standardization of molecular biology experiments such as RT-PCR and Western blot, especially It is used as an internal standard protein in the relative expression quantification of proteins in tissues or cells. Although GAPDH has been widely used as an internal standard protein, previous studies have only focused on relative quantification of protein expression in eukaryotic organisms.
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[4]Glyceraldehyde-3-phosphate dehydrogenase:A universal internal control for Western blots in prokaryotic and eukaryotic cells[J].Yonghong Wu,Mi Antioxidant 168n Wu,Guowei He,Xiao Zhang, Weiguang Li,Yan Gao,Zhihui Li,Zhaoyan Wang,Chenggang Zhang.Analytical Biochemistry.2012(1)
[5]Wu Yonghong.GAPDH is a universal drug for prokaryotic and eukaryotic organisms.��Research on internal standard proteins and related biotechnology research and development[D]. Academy of Military Medical Sciences of the Chinese People’s Liberation Army, 2013.
