Key features and details
- Assay type: Enzyme activity (quantitative)
- Detection method: Colorimetric/Fluorometric
- Platform: Microplate reader
- Assay time: 40 min
- Sample type: Cell culture supernatant, Other biological fluids, Plasma, Serum, Tissue, Urine
- Sensitivity: 0.1 mU/ml
Product namePyruvate Kinase Assay Kit
Sample typeCell culture supernatant, Urine, Serum, Plasma, Other biological fluids, Tissue
Assay typeEnzyme activity (quantitative)
Sensitivity= 0.1 mU/ml
Assay time0h 40m
Pyruvate Kinase Assay Kit ab83432 provides a simple, direct and automation-ready assay 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 pyruvate kinase assay protocol, PEP and ADP are catalyzed by pyruvate kinase to generate pyruvate and ATP. The generated pyruvate is oxidized by pyruvate oxidase to produce color (absorbance 570 nm) and fluorescence (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.
Pyruvate kinase assay protocol summary:
- add samples and standards to wells
- add reaction mix
- analyze with microplate reader immediately and after 10-20 min
This product is manufactured by BioVision, an Abcam company and was previously called K709 Pyruvate Kinase Activity Colorimetric/Fluorometric Assay Kit. K709-100 is the same size as the 100 test size of ab83432.
Pyruvate kinase (PK, EC 22.214.171.124) 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.
Storage instructionsStore at -20°C. Please refer to protocols.
Components Identifier 100 tests 2000 tests OxiRed Probe Red 1 x 200µl 20 x 200µl PK Assay Buffer WM 1 x 25ml 20 x 25ml PK Enzyme Mix (Lyophilised) Green 1 vial 20 vials PK Positive Control (Lyophilised) Blue 1 vial 20 vials PK Substrate Mix (Lyophilised) Purple 1 vial 20 vials Pyruvate Standard (100 nmol/µl) Yellow 1 x 100µl 20 x 100µl
RelevancePyruvate 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.
Cellular localizationCytoplasmic and Nuclear
- EC 188.8.131.52
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.
Pyruvate measured in biological fluid after 2 min, 10 min and 20 min incubation time showing quantity (nmol) per ml of tested sample.
Pyruvate measured in mouse tissue lysates after 2 min, 10 min and 20 min incubation time showing quantity (nmol) per mg of tested sample.
Pyruvate measured in cell lysates after 2 min, 10 min and 20 min incubation time showing quantity (nmol) per 1 mln of tested cells.
Colorimetric standard curve: mean of duplicates (+/- SD) with background reads subtracted.
Fluorometric standard curve: mean of duplicates (+/- SD) with background reads subtracted.
Pyruvate kinase samples time line using ab83432.
Pyruvate standard curve using ab83432.
ab83432 has been referenced in 22 publications.
- Li X et al. Discovery of Functional Alternatively Spliced PKM Transcripts in Human Cancers. Cancers (Basel) 13:N/A (2021). PubMed: 33478099
- Li Y et al. Hedgehog interacting protein (HHIP) represses airway remodeling and metabolic reprogramming in COPD-derived airway smooth muscle cells. Sci Rep 11:9074 (2021). PubMed: 33907231
- Ji L et al. Worenine reverses the Warburg effect and inhibits colon cancer cell growth by negatively regulating HIF-1a. Cell Mol Biol Lett 26:19 (2021). PubMed: 34006215
- Na AY et al. Global Lysine Acetylome Analysis of LPS-Stimulated HepG2 Cells Identified Hyperacetylation of PKM2 as a Metabolic Regulator in Sepsis. Int J Mol Sci 22:N/A (2021). PubMed: 34445236
- Cheng J et al. TRIM21 and PHLDA3 negatively regulate the crosstalk between the PI3K/AKT pathway and PPP metabolism. Nat Commun 11:1880 (2020). PubMed: 32312982