EnzyChrom™ Creatine Kinase Assay Kit
Datasheet PDF  MSDS/Bulk Order


Application
For quantitative determination of creatine kinase activity and evaluation of drug effects on CK activity.

Key Features
Sensitive and accurate. Detection range: 5 to 300 U/L creatine kinase in 96-well plate assay.

Convenient. The procedure involves adding a single working reagent, and reading the optical density at 20 min and 40 min at room temperature or 37°C.

High-throughput. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day.

Method
OD340nm

Samples
Serum, plasma etc

Species
All

Procedure
5 min

Size
100 tests

Detection Limit
5 U/L

Shelf Life
6 months

More Details
CREATINE KINASE (CK), also known as creatine phosphokinase (CPK), is an enzyme (EC 2.7.3.2) expressed predominantly in skeletal muscle, smooth muscle and the brain. The CK enzyme consists of two subunits, which can be either B (brain type) or M (muscle type), and hence three different isoenzymes: CK-MM, CK-BB and CK-MB. CK catalyzes the conversion of creatine to phosphocreatine, consuming adenosine triphosphate (ATP) and generating adenosine diphosphate (ADP) and the reverse reaction. CK is often determined routinely in emergency patients with chest pain and acute renal failure. Elevation of CK is an indication of damage to muscle and has been associated with injury, rhabdomyolysis, myocardial infarction, myositis, myocarditis, malignant hyperthermia and neuroleptic malignant syndrome, etc. Lower levels can be an indication of alcoholic liver disease and rheumatoid arthritis. Simple, direct and automation-ready procedures for measuring CK activity are very desirable. BioAssay Systems’ EnzyChrom™ Creatine Kinase Assay Kit is based on enzyme coupled reactions in which creatine phosphate and ADP is converted to creatine and ATP by CK, the generated ATP is used to phosphorylate glucose by hexokinase to generate glucose-6-phosphate, which is then oxidized by NADP in the presence of glucose-6-phosphate dehydrogenase. The produced NADPH, measured at 340 nm, is proportionate to the CK activity in the sample.

1. Our instrument does not have a filter for 340 nm. Can I measure at 360 nm instead?



No, unfortunately, this is not possible, because the calibrator does not absorb at 360 nm. The assay must be read at 340 nm.

2. I am using the EnzyChrom Creatine Kinase kit (ECPK-100) but my serum samples are hemolysed. What can I do to avoid interference in the assay?



Hemolysis releases adenylate kinase from red blood cells into the blood, which causes an apparent increase in CPK activity. The best solution would be to collect fresh samples and try to avoid hemolysis.

3. How to prepare tissue homogenates for use in the assay?



For tissue sample preparation, we recommend the following:

1. Homogenize tissue in phosphate buffered saline. This can be done using a homogenizer (e.g. the Dounce type) or a grinder. If you use a lysis buffer, the buffer should not contain ascorbic acid, SDS, sodium azide, NP-40 and Tween 20.

2. Centrifuge to pellet any debris. This can be done at room temperature in a table centrifuge (e.g. 5 min at 14,000 rpm). The supernatant should be clear.

3. Use the supernatant for CPK assays.

For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions
1. Kim, YM et al (2009). Palmatine from Coptidis rhizoma reduces ischemia-reperfusion-mediated acute myocardial injury in the rat. Food Chem Toxicol 47(8):2097-102. Assay: Creatine Kinase in Rat Serum (Pubmed).

2. Branciari, R et al (2009). Effect of genotype and rearing system on chicken behavior and muscle fiber characteristics. J Anim Sci 87(12):4109-17. Assay: Creatine Kinase in Chicken Serum (Pubmed).

3. Annecke, T et al (2010). Sevoflurane preserves the endothelial glycocalyx against ischaemia-reperfusion injury. Br J Anaesth 104(4):414-21. Assay: Creatine Kinase in Guinea Pig Heart transudate (Pubmed).

4. Kwon, S et al (2010). ASB9 interacts with ubiquitous mitochondrial creatine kinase and inhibits mitochondrial function. BMC Biol 8:23. Assay: Creatine Kinase in Human HEK 293 cells (Pubmed).

5. Steen, C et al (2010). Reduced creatine kinase B activity in multiple sclerosis normal appearing white matter. PLoS One 5(5):e10811. Assay: Creatine Kinase in Human Brain tissue (Pubmed).

6. Pruksakorn, D et al (2010). Metabolic alteration of HepG2 in scaffold-based 3-D culture: proteomic approach. Proteomics 10(21):3896-904. Assay: Creatine Kinase in Human HepG2 hepatocellular carcinoma cells (Pubmed).

7. Lu, X et al (2009). Activation of NF-kappaB is a critical element in the antiapoptotic effect of anesthetic preconditioning. Am J Physiol Heart Circ Physiol 296(5):H1296-304. Assay: Creatine Kinase in Rat Heart perfusate (Pubmed).

8. Itaka, K et al (2010). Polyplex nanomicelle promotes hydrodynamic gene introduction to skeletal muscle. J Control Release 143(1):112-9. Assay: Creatine Kinase in Mouse Serum (Pubmed).

9. Shi, H et al (2010). MAP kinase phosphatase-1 deficiency impairs skeletal muscle regeneration and exacerbates muscular dystrophy. FASEB J 24(8):2985-97. Assay: Creatine Kinase in Mouse Serum (Pubmed).

10. Gawlik KI, Durbeej M (2010). Transgenic overexpression of laminin alpha1 chain in laminin alpha2 chain-deficient mice rescues the disease throughout the lifespan. Muscle Nerve 42(1):30-7. Assay: Creatine Kinase in Mouse Serum (Pubmed).

11. Lu, X et al (2011). Phosphorylation of ARC is a critical element in the antiapoptotic effect of anesthetic preconditioning. Anesth Analg 112(3):525-31. Assay: Creatine Kinase in Rat Heart perfusate (Pubmed).

12. Li, S et al (2011). Reduction of cold ischemia-reperfusion injury by graft-expressing clusterin in heart transplantation. J Heart Lung Transplant 30(7):819-26. Assay: Creatine Kinase in Mouse Heart preservation solution (Pubmed).

13. Ren, JY et al (2011). Cardioprotection by ischemic postconditioning is lost in isolated perfused heart from diabetic rats: Involvement of transient receptor potential vanilloid 1, calcitonin gene-related peptide and substance P. Regul Pept 169(1-3):49-57. Assay: Creatine Kinase in Rat Heart perfusate (Pubmed).

14. Uchida, S et al (2011). Combination of chondroitin sulfate and polyplex micelles from Poly(ethylene glycol)-poly{N’-[N-(2-aminoethyl)-2-aminoethyl]aspartamide} block copolymer for prolonged in vivo gene transfection with reduced toxicity. J Control Release . Assay: Creatine Kinase in Mouse Serum (Pubmed).

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EnzyChrom™ Creatine Kinase Assay Kit
Catalog No: ECPK-100
Price: $399    Qty:
For orders of 10 or more kits, please contact us at 1-510-782-9988 x 1 for best pricing.

Shipping: On Ice
Shipment: Same Day for order by 2pm PST
Delivery: 1-2 days (US), 3-6 days (Intl);
Storage: -20°C



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