【专题讨论】如何选择内参？
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关于内参，已经有很多的帖子讨论，但是在我们的动物或细胞的模型中，总是有各种干扰的过程常常影响内参的表达，比如GAPDH就参与糖代谢，变性性疾病，凋亡等多种过程，如果我们的试验中存在这种过程干扰内参的表达，应该如何选择呢？Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody
Glyseraldehyde-3-phosphate dehydrogenase (GAPDH) is a catalytic enzyme involved in glycolysis. Besides its functioning as a glycolytic enzyme within the cytoplasm, GAPDH is also involved in early steps of apoptosis. Recent evidence suggests that mammalian GAPDH is also involved in membrane fusion, microtubule bundling, phosphotransferase activity, nuclear RNA export, prostate cancer progression, programmed neuronal cell death, DNA replication, and DNA repair. GAPDH is constitutively expressed in almost all tissues at high levels. There are however some physiological factors, such as hypoxia and diabetes, that increase GAPDH expression in certain cell types. GAPDH may also be involved in age-related neuronal diseases, such as Alzheimer’s and Huntington’s disease.
Monoclonal antibodies to GAPDH
GAPDH is composed of four 30-40 kDa subunits. The molecular weight for GAPDH is 146 kDa. Since it is expressed in all cells it is for example becoming the marker of choice for a loading control in Western Blotting. The anti-GAPDH clone 6C5, which is developed at HyTest Ltd, gives very strong band in Western Blotting at dilution 1/2000, but it can be used down to 1/10,000 dilution.
Glyseraldehyde-3-phosphate dehydrogenase (GAPDH) is a catalytic enzyme involved in glycolysis. Besides its functioning as a glycolytic enzyme within the cytoplasm, GAPDH is also involved in early steps of apoptosis. Recent evidence suggests that mammalian GAPDH is also involved in membrane fusion, microtubule bundling, phosphotransferase activity, nuclear RNA export, prostate cancer progression, programmed neuronal cell death, DNA replication, and DNA repair. GAPDH is constitutively expressed in almost all tissues at high levels. There are however some physiological factors, such as hypoxia and diabetes, that increase GAPDH expression in certain cell types. GAPDH may also be involved in age-related neuronal diseases, such as Alzheimer’s and Huntington’s disease.
Monoclonal antibodies to GAPDH
GAPDH is composed of four 30-40 kDa subunits. The molecular weight for GAPDH is 146 kDa. Since it is expressed in all cells it is for example becoming the marker of choice for a loading control in Western Blotting. The anti-GAPDH clone 6C5, which is developed at HyTest Ltd, gives very strong band in Western Blotting at dilution 1/2000, but it can be used down to 1/10,000 dilution.
