| Product Includes | Product # | Quantity | Color | Storage Temp |
|---|---|---|---|---|
| Phospho-Pyruvate Dehydrogenase α1 (Ser293) Rabbit mAb Coated Microwells | 30221 | 96 tests |
|
+4C |
| Pyruvate Dehydrogenase α1 Rabbit Detection mAb | 52531 | 1 ea |
|
+4C |
| HRP Diluent | 13515 | 5.5 ml |
|
+4C |
| TMB Substrate | 7004 | 11 ml |
|
+4C |
| STOP Solution | 7002 | 11 ml |
|
+4C |
| Sealing Tape | 54503 | 2 ea |
|
+4C |
| ELISA Wash Buffer (20X) | 9801 | 25 ml |
|
+4C |
| Cell Lysis Buffer (10X) | 9803 | 15 ml |
|
-20C |
*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.
Description
The rapid protocol (RP) PathScan® RP Phospho-Pyruvate Dehydrogenase α1 (Ser293) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of pyruvate dehydrogenase α1 protein phosphorylated at Ser293 in a reduced assay time of 1.5 hours. Incubation of cell lysates and detection antibody on the coated microwell plate forms a sandwich with phospho-pyruvate dehydrogenase α1 (Ser293) in a single step. The plate is then extensively washed and TMB reagent is added for signal development. The magnitude of absorbance for the developed color is proportional to the quantity of pyruvate dehydrogenase α1 protein phosphorylated at Ser293. Learn more about your ELISA kit options here.
*Antibodies in this kit are custom formulations specific to kit.
Specificity/Sensitivity
Background
The pyruvate dehydrogenase complex catalyzes the conversion of pyruvate and CoA into acetyl-CoA and CO2 in the presence of NAD+. Acetyl-CoA then goes into the citric acid cycle where it reacts with oxaloacetate to form citrate. The reaction of oxidative decarboxylation of pyruvate serves as a critical link between glycolysis and the citric acid cycle. In mammalian cells, the pyruvate dehydrogenase complex is located in the mitochondrial matrix (1). This complex is composed of three enzymes: pyruvate dehydrogenase (E1), dihydrolipoamide acetyltransferase (E2), and dihydrolipoamide dehydrogenase (E3). Pyruvate dehydrogenase (E1) consists of two subunits: α and β. This enzyme catalyzes the removal of CO2 from pyruvate. Mutations in the α subunits of pyruvate dehydrogenase (E1) lead to congenital defects that are usually associated with lactic acidosis, neurodegeneration, and early death (2).
Pyruvate dehydrogenase kinase 1 phosphorylates pyruvate dehydrogenase (E1) α1 subunit at Ser293 to inactivate its activity (3,4). This phosphorylation contributes to the tumor metabolic reprogramming toward glycolysis in hypoxia by inhibiting the citric acid cycle (4).
Background References
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
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