EnzyChrom™ Creatine Kinase Assay Kit
Application
- For quantitative determination of creatine kinase (CK) 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
- 40 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.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.
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.
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.
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.
Yoon, JA et al. (2020). Evaluation of changes in body composition and proinflammatory marker levels in ovariectomized rats with induced hyperthyroidism. All Life, 13(1): 99-107. Assay: Creatine Kinase in rat serum.
Starkoff, B et al. (2020. Protein Supplementation Does Not Enhance Recovery from Exercise-Induced Muscle Damage. Journal of Exercise Physiology Online, 23(1): 99-112. Assay: Creatine Kinase in human serum.
Yang, JH et al. (2020). Interaction of OIP5-AS1 with MEF2C mRNA promotes myogenic gene expression. Nucleic Acids Research, 48(22): 12943-12956. Assay: Creatine Kinase in mouse and human cells.
Kodsi, DNE et al. (2020). Parkinson disease-linked parkin mediates redox reactions that lower oxidative stress in mammalian brain. BioRxiv, 2020.04.26.062380. Assay: Creatine Kinase in mouse mitochondria.
Melendez, DM et al. (2020). Effect of transport and rest stop duration on the welfare of conditioned cattle transported by road. PloS One, 15(3), e0228492. Assay: Creatine Kinase in bovine serum.
Melendez, DM et al. (2021). Effects of conditioning, source, and rest on indicators of stress in beef cattle transported by road. PloS One, 16(1), e0244854. Assay: Creatine Kinase in bovine serum.
Ramani Sattiraju, S et al. (2020). Loss of membrane integrity drives myofiber death in lipin1-deficient skeletal muscle. Physiological Reports, 8(20). Assay: Creatine Kinase in mouse serum.
Kitazume-Taneike, Rika, et al (2019). Ablation of Toll-like receptor 9 attenuates myocardial ischemia/reperfusion injury in mice. Biochemical and Biophysical Research Communications 515(3):442-447. Assay: Creatine kinase in mice coronary effluent.
Munk, Rachel, et al (2019). Loss of miR-451a enhances SPARC production during myogenesis. PloS one 14.3: e0214301. Assay: Creatine kinase in mouse cells.
Zelikovich, Aaron S., et al (2019). Moderate exercise improves function and increases adiponectin in the mdx mouse model of muscular dystrophy. Scientific reports 9.1: 5770. Assay: Creatine kinase in mouse serum.
Demoy-Schneider, Marina, et al (2018). Quality assessment of cryopreserved black-lip pearl oyster Pinctada margaritifera spermatozoa. Aquaculture 497: 278-286. Assay: Creatine kinase in oyster tissue.
Horn, Adam (2018). Role of Mitochondria in Plasma Membrane Repair and Pathogenesis of MuscularDystrophy. Diss. The George Washington University. Assay: Creatine kinase in mice serum.
McNally, Elizabeth (2018). Antisense Polynucleotides to Induce Exon Skipping and Method of Treating Dystrophies. U.S. Patent Application No. 16/041,278. Assay: Creatine kinase in mice serum.
Dowman, Leona M., et al (2017). Greater endurance capacity and improved dyspnoea with acute oxygen supplementation in idiopathic pulmonary fibrosis patients without resting hypoxaemia. Respirology 22.5: 957-964. Assay: Creatine kinase in human plasma.
Mattioli, S., et al (2017). Adaptive response to exercise of fast-growing and slow-growing chicken strains: Blood oxidative status and non-enzymatic antioxidant defense. Poultry science 96.11: 4096-4102. Assay: Creatine kinase in chicken serum.
Quattrocelli, Mattia, et al (2017). Genetic modifiers of muscular dystrophy act on sarcolemmal resealing and recovery from injury. PLoS genetics 13.10: e1007070. Assay: Creatine kinase in mouse serum.
Quattrocelli, Mattia, et al (2017). Intermittent glucocorticoid dosing improves muscle repair and function in mice with limb-girdle muscular dystrophy. The American journal of pathology 187.11: 2520-2535. Assay: Creatine kinase in mouse serum.
Weis, Sebastian, et al (2017). Metabolic adaptation establishes disease tolerance to sepsis. Cell 169.7: 1263-1275. Assay: Creatine kinase in mice plasma.
Chali, Farah, et al (2016). Long-term exercise-specific neuroprotection in spinal muscular atrophy-like mice. The Journal of physiology 594.7: 1931-1952. Assay: Creatine kinase in mice serum.
Demonbreun, Alexis R., et al (2016). Enhanced muscular dystrophy from loss of dysferlin is accompanied by impaired annexin A6 translocation after sarcolemmal disruption. The American journal of pathology 186.6: 1610-1622. Assay: Creatine kinase in mice plasma.
Ju, Huiming, et al (2016). MicroRNA-378 promotes myogenic differentiation by targeting BMP4. Molecular medicine reports 13.3: 2194-2200. Assay: Creatine kinase in mouse cells.
Panda, Amaresh C., et al (2016). Novel RNA-binding activity of MYF5 enhances Ccnd1/Cyclin D1 mRNA translation during myogenesis. Nucleic acids research 44.5: 2393-2408. Assay: Creatine kinase in mouse cells.
Park, Sang Chul, et al (2016). Pharmaceutical composition for preventing or treating muscle wasting-related disease comprising diaminodiphenylsulfone or pharmaceutically acceptable salt thereof. U.S. Patent No. 9,226,905. Assay: Creatine kinase in mice blood.
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.
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.
Ren, J.Y., 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.
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 155(2):296-302. Assay: Creatine kinase in mouse serum.
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.
Gawlik, K.I.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.
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.
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 cells.
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 carcinoma cells.
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.
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.
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.
Kim, Y.M., 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.
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.
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If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.
– Fast turnaround
– Quality data
– Low cost