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Pyruvate Kinase (PK) Assay Kit (ab83432) provides a simple, direct and automation-ready procedure for measuring pyruvate kinase activity in cell culture supernatant, plasma, serum, tissue, urine and other biological fluids (not validated for use with whole blood). In the assay, PEP and ADP were catalyzed by PK to generate pyruvate and ATP. The generated pyruvate is oxidized by pyruvate oxidase to produce color (at λ = 570 nm) and fluorescence (at Ex/Em = 535/587 nm). Since the increase in color or fluorescence intensity is proportional to the increase in pyruvate amount, the PK activity can be accurately measured. The kit detects 0.1 mU/ml pyruvate kinase.
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Pyruvate kinase (PK, EC 126.96.36.199) is an enzyme involved in glycolysis. It catalyzes the transfer of a phosphate group from phosphoenolpyruvate (PEP) to ADP, yielding one molecule of pyruvate and one molecule of ATP. Lack of pyruvate kinase will slow down the process of glycolysis which causes the disease known as pyruvate kinase deficiency.
|OxiRed Probe||Red||1 x 200µl|
|PK Assay Buffer||WM||1 x 25ml|
|PK Enzyme Mix (Lyophilised)||Green||1 vial|
|PK Positive Control (Lyophilised)||Blue||1 vial|
|PK Substrate Mix (Lyophilised)||Purple||1 vial|
|Pyruvate Standard (100 nmol/µl)||Yellow||1 x 100µl|
Our Abpromise guarantee covers the use of ab83432 in the following tested applications.
|Functional Studies||Use at an assay dependent dilution.|
Pyruvate measured in biological fluid after 2 min, 10 min and 20 min incubation time showing quantity (nmol) per ml of tested sample.
Fluorometric standard curve: mean of duplicates (+/- SD) with background reads subtracted.
PC-3 and MCF-7 cells were treated with OA at the indicated doses and PK activity was determined 12 hr after OA treatment using ab83432. The average values of three independent experiments were shown as Mean ± SD. *, P<0.05, **, P<0.01.