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MAK116

Sigma-Aldrich

Creatine Kinase Activity Assay Kit

sufficient for 100 tests (Ultraviolet)

Synonym(s):

CK Activity Assay Kit, Creatine Phosphokinase Activity Assay Kit

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About This Item

UNSPSC Code:
12161503
NACRES:
NA.84
Pricing and availability is not currently available.

usage

sufficient for 100 tests (Ultraviolet)

detection method

colorimetric

relevant disease(s)

cardiovascular diseases; genitourinary disorders; aging/geriatric diseases; orthopedic diseases

shipped in

dry ice

storage temp.

−20°C

Gene Information

human ... CKB(1152) , CKM(1158)
mouse ... CKB(12709) , CKM(12715)
rat ... CKB(24264) , CKM(24265)

Related Categories

General description

Creatine Kinase (CK), also known as phosphocreatine kinase, is an enzyme that catalyzes the transfer of one phosphate group from ATP to creatine generating phosphocreatine, an important energy reservoir in muscle and brain tissue. CK is a dimeric protein made up of B (brain) and M (muscle) subunits. Three isoenzymes, CK-MM, CK-MB, and CK-BB, have been observed. CK levels are elevated in various pathological conditions including myocardial infarction, rhabdomyolysis, muscular dystrophy, and renal failure.

Features and Benefits

Compatible with high-throughput handling systems.

Suitability

Suitable for the detection of Creatine Kinase activity in a variety of samples such as serum and plasma. This kit can be used to study the effects of various compounds on Creatine Kinase activity.

Principle

Creatine Kinase activity is determined by a coupled enzyme reaction resulting in the production of NADPH, measured at 340 nm, proportionate to the CK activity present in the sample. In this reaction, phosphocreatine and ADP are converted to creatine and ATP. The generated ATP is used by hexokinase to phosphorylate glucose resulting in glucose-6-phosphate, which is oxidized by NADP in the presence of glucose-6-phosphate dehydrogenase to produce NADPH and 6-phospho-D-gluconate. One unit of CK is the amount of enzyme that will transfer 1.0 μmole of phosphate from phosphocreatine to ADP per minute at pH 6.0. This kit has a linear range of 30–1,800 units/L CK activity.

related product

Product No.
Description
Pricing

Pictograms

Health hazard
GHS08

Signal Word

Danger

Hazard Statements

H334

Precautionary Statements

P261 - P284 - P501

Hazard Classifications

Resp. Sens. 1

Storage Class Code

12 - Non Combustible Liquids

Choose from one of the most recent versions:

Certificates of Analysis (COA)

Lot/Batch Number
116CE12A18
116CE12A18
116CE09A26
116CE10A11
116CE09A26

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Marcus Waskiw-Ford et al.
Nutrients, 12(4) (2020-04-16)
Leucine-enriched essential amino acids (LEAAs) acutely enhance post-exercise myofibrillar protein synthesis (MyoPS), which has been suggested to be important for muscle repair and recovery. However, the ability of LEAAs to concurrently enhance MyoPS and muscle damage recovery in free-living humans
Evaluation of White Sesame Seed Oil on Glucose Control and Biomarkers of Hepatic, Cardiac, and Renal Functions in Male Sprague-Dawley Rats with Chemically Induced Diabetes
Aslam F, et al.
Journal of Medicinal Food, 448-57 null
HANAC Col4a1 Mutation in Mice Leads to Skeletal Muscle Alterations due to a Primary Vascular Defect
Guiraud S, et al.
The American Journal of Pathology, 505-516 null
Gary R Hunter et al.
Medicine and science in sports and exercise, 49(12), 2414-2421 (2017-07-25)
Resting energy expenditure (REE) increases after an intense exercise; however, little is known concerning mechanisms. The purpose of this study was to determine effects of a single bout of moderate-intensity continuous (MIC) aerobic exercise, or high-intensity interval (HII) exercise on
P2RX7 Purinoceptor: A Therapeutic Target for Ameliorating the Symptoms of Duchenne Muscular Dystrophy
Sinadinos A
PLoS Medicine null
View All Related Papers

Questions

1–4 of 4 Questions  

  1. How long are the provided solutions stable for when stored at -20C? Does the assay lose efficacy through repeated freeze/thaw cycles of the assay solutions?

    1 answer

    1. The unopened kit is expiration dated on a batch-by-batch basis and the certificate of analysis for a specific lot would need to be reviewed for this information. Please review the "product documentation" section of the website to pull a sample CoA or to input the lot number for the received lot.

      https://www.sigmaaldrich.com/product/sigma/MAK116#product-documentation

      There is no data available on the amount of freeze/thaws this kit can withstand. It is expected that repeated freeze/thaws of enzyme and substrate solution could affect the activity. Therefore, it would be advised to aliquot multiple small volumes of the kit reagents for one-time use to avoid repeated freeze/thaw cycles.

      Helpful?

  2. What is the required cell count for this procedure, and at what cell concentration is the supernatant obtained?

    1 answer

    1. For preparing cell samples for use with MAK116, the Creatine Kinase Activity Assay Kit, the vendor recommends using the cell lysate for the assay. The number of cells to use will depend on the CK activity of the cells, and the final CK activity of the sample used for the assay should be within 5 to 300 U/L creatine kinase (the linear detection range in a 96-well plate assay). Here's an example of a sample preparation protocol for adherent or suspension cells:

      1. Harvest the required number of cells for each assay, with an initial recommendation of two million or 2 x 10^6 cells.
      2. Wash the cells with cold PBS and resuspend them in 400 µL PBS on ice.
      3. Lyse the cells by homogenization (10-20 passes in a Dounce homogenizer on ice) or by sonication (preferably performed in an ice-water bath). The degree of cell lysis can be checked under a microscope.
      4. Centrifuge the homogenate at 14,000 g for 10 minutes and transfer the clear supernatant into a clean tube.
      5. Conduct a pilot test of the sample at different dilutions. Choose a dilution with readings within the range of the calibrator for further assays. Most samples can be stored at -80°C if not assayed immediately.

      Helpful?

  3. Can this kit be used on an adherent cell line (rat species)?

    1 answer

    1. Creatine kinase is generally found in muscle tissue or in plasma. The MAK116 has been published in the scientific literature for samples such as rat serum, plasma and tissue. There are not any articles referring specifically to rat adherent cell lines. Further research and development for this kit would be necessary to validate for this application. The insert does provide instructions for working with adherent cell lysate.
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/355/268/mak116bul.pdf

      Helpful?

  4. Regarding the creatine kinase assay, will this detect CK activity from other mammalian species such as bovine?

    1 answer

    1. Yes, this kit is suitable for determining CK levels in mammalian samples.

      Helpful?

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