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Pyruvate Kinase Activity Colorimetric/Fluorometric Assay Kit

based on 7 citations in multiple journalsPyruvate Kinase Activity Colorimetric/Fluorometric Assay Kit74.2 4
Sensitive Assay, HTS
Catalog #: K709

Product Details

Cat # +Size K709-100
Size 100 assays
Detection Method Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm)
Species Reactivity Mammalian
Applications The kit detects 0.1 mU/ml pyruvate kinase.
Features & Benefits • Simple procedure; takes ~ less than 40 minutes
• Fast and convenient
Kit Components • PK Assay Buffer
• OxiRed™ Probe (in DMSO)
• PK Enzyme Mix
• PK Substrate Mix
• PK Positive Control (~18 mU)
• Pyruvate Standard (100 nmol/µl)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.


Pyruvate kinase (PK, EC 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. BioVision provides a simple, direct and automation-ready procedure for measuring pyruvate kinase activity in various biological samples such as blood, tissues, and culture cells, etc. 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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The PK kit has no instructions for how to homogenize tissues for the kit. Do you know if I can use the same assay buffer to make lysates to run both assays? How much muscle tissue is recommended?
Here’s a general protocol to preparing tissue lysates: Start with 10-20 mg of the muscle tissue, add 200-500 µl (or ~4-6 volumes) of the assay buffer on ice, homogenize using a Dounce homogenizer (10-50 passes) on ice, until efficient lysis is confirmed, by viewing the cells under the microscope. Spin down the sample and collect the supernatant. Use the supernatant for your subsequent assays. Appropriate dilutions of the sample must be tested in order ensure the readings will fall within the linear range of the standard curve.
How long is the reaction mix (See table 1) stable for? Can I make up a stock of the reaction mix and use it throughout the day? Do I have to wait for a stable colour to be achieved for the reaction mix or can I add to sample straight away?
We don’t recommend storing the reaction mix for a long time. The enzymes are susceptible to degradation. Plus multiple usage of the same vial can lead to contamination and can cause interference in the reaciton.
Can we use frozen samples with this assay?
We would recommend using fresh samples. However, frozen samples can be used too provided they have not undergone multiple freeze thaw cycles.
Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
What is the shelf life of this kit?
BioVisions’s assay kits expire 6-12 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet.
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Dhanush Haspula, Influence of a Hyperglycemic Microenvironment on a Diabetic Versus Healthy Rat Vascular Endothelium Reveals Distinguishable Mechanistic and Phenotypic Responses. Front Physio, Feb 2019;  31133884.
Lu CY et al., (2017) Aging results in a decline in cellular energy metabolism in the trophocytes and oenocytes of worker honeybees (Apis mellifera), Adipologie, 2017, June 23: epub
Park, Seong-Hoon et al. (2016) SIRT2-Mediated Deacetylation and Tetramerization of Pyruvate Kinase Directs Glycolysis and Tumor Growth. Cancer Res. 2016 Jul 1;76(13):3802-12.
Nakatsu et al., L-cysteine reversibly inhibits glucose-induced biphasic insulin secretion and ATP production by inactivating PKM2. PNAS, Mar 2015; 112: E1067 - E1076.
Park et al., Changes in Pyruvate Metabolism Detected by Magnetic Resonance Imaging Are Linked to DNA Damage and Serve as a Sensor of Temozolomide Response in Glioblastoma Cells.Cancer Res., Dec 2014; 74: 7115 - 7124.
For more citations of this product click here