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| Sample Type(s) | Serum, plasma etc |
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Overview
For quantitative determination of creatine kinase (CK) activity and evaluation of drug effects on CK activity. The assay uses OD340nm for signal readout. Compatible sample input includes Serum, plasma etc. Typical stated assay timing is 40 min.
Key elements and design rationale
- Readout format: OD340nm supports plate-based signal acquisition and consistent comparison across matched samples.
- Sample compatibility: The stated sample scope includes Serum, plasma etc, which is useful when aligning matrix type with calibration and control design.
- Analytical range context: The supplied specifications include a stated detection limit of 5 U/L for interpreting low-signal samples.
- Feature emphasis: Sensitive and accurate. Detection range: 5 to 300 U/L creatine kinase in 96-well plate assay.
Additional feature notes highlight 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. Available format information for this listing includes 100 Tests.
Biological background
This product is centered on measurement of creatine kinase within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.
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.
Detection method
Colorimetric (OD 340 nm).
Detection limit and analytical sensitivity
Reported detection limit: 5 U/L.
Procedures and timing
Stated procedure or timing information: 40 min.
Research relevance and current trends
- Plate-based quantification and side-by-side group comparison remain central use cases for this assay format.
- The product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.
- The description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.
Common research applications
- Quantify creatine kinase in serum, plasma by OD340 nm readout.
- Compare treatment or phenotype groups using matched serum, plasma handling.
- Monitor time-course or pre/post changes in serum, plasma across study conditions.
Interpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.
Notes for experimental interpretation
- Matrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.
- Use appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.
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.
For laboratories requiring additional technical capacity, we provide scientific support services including assay execution, method guidance, product sourcing, and customization to align the assay with specific experimental objectives. If you need assistance selecting the appropriate kit configuration, adapting the workflow to your application, or identifying related research services, please click Talk to a Scientist, email support@biohippo.com, or review our Research Services; a member of our scientific team will follow up with recommendations tailored to your study.
Evaluation of changes in body composition and proinflammatory marker levels in ovariectomized rats with induced hyperthyroidism
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
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.
Interaction of OIP5-AS1 with MEF2C mRNA promotes myogenic gene expression
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.
Parkinson disease-linked parkin mediates redox reactions that lower oxidative stress in mammalian brain
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.
Effect of transport and rest stop duration on the welfare of conditioned cattle transported by road
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.
Effects of conditioning, source, and rest on indicators of stress in beef cattle transported by road
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.
Loss of membrane integrity drives myofiber death in lipin1-deficient skeletal muscle
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.
Ablation of Toll-like receptor 9 attenuates myocardial ischemia/reperfusion injury in mice
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.
Loss of miR-451a enhances SPARC production during myogenesis
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.
Moderate exercise improves function and increases adiponectin in the mdx mouse model of muscular dystrophy
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.
Quality assessment of cryopreserved black-lip pearl oyster Pinctada margaritifera spermatozoa
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.
Role of Mitochondria in Plasma Membrane Repair and Pathogenesis of MuscularDystrophy
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.
Antisense Polynucleotides to Induce Exon Skipping and Method of Treating Dystrophies
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.
Greater endurance capacity and improved dyspnoea with acute oxygen supplementation in idiopathic pulmonary fibrosis patients without resting hypoxaemia
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.
Adaptive response to exercise of fast-growing and slow-growing chicken strains: Blood oxidative status and non-enzymatic antioxidant defense
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.
Genetic modifiers of muscular dystrophy act on sarcolemmal resealing and recovery from injury
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.
Intermittent glucocorticoid dosing improves muscle repair and function in mice with limb-girdle muscular dystrophy
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.
Metabolic adaptation establishes disease tolerance to sepsis
Weis, Sebastian, et al (2017). Metabolic adaptation establishes disease tolerance to sepsis. Cell 169.7: 1263-1275. Assay: Creatine kinase in mice plasma.
Long-term exercise-specific neuroprotection in spinal muscular atrophy-like mice
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.
Enhanced muscular dystrophy from loss of dysferlin is accompanied by impaired annexin A6 translocation after sarcolemmal disruption
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.
MicroRNA-378 promotes myogenic differentiation by targeting BMP4
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.
Novel RNA-binding activity of MYF5 enhances Ccnd1/Cyclin D1 mRNA translation during myogenesis
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.
Pharmaceutical composition for preventing or treating muscle wasting-related disease comprising diaminodiphenylsulfone or pharmaceutically acceptable salt thereof
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.
Reduction of cold ischemia-reperfusion injury by graft-expressing clusterin in heart transplantation
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.
Phosphorylation of ARC is a critical element in the antiapoptotic effect of anesthetic preconditioning
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.
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
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.
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
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.
Sevoflurane preserves the endothelial glycocalyx against ischaemia-reperfusion injury
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.
Transgenic overexpression of laminin alpha1 chain in laminin alpha2 chain-deficient mice rescues the disease throughout the lifespan
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.
Polyplex nanomicelle promotes hydrodynamic gene introduction to skeletal muscle
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.
ASB9 interacts with ubiquitous mitochondrial creatine kinase and inhibits mitochondrial function
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.
Metabolic alteration of HepG2 in scaffold-based 3-D culture: proteomic approach
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.
MAP kinase phosphatase-1 deficiency impairs skeletal muscle regeneration and exacerbates muscular dystrophy
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.
Reduced creatine kinase B activity in multiple sclerosis normal appearing white matter
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.
Effect of genotype and rearing system on chicken behavior and muscle fiber characteristics
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.
Palmatine from Coptidis rhizoma reduces ischemia-reperfusion-mediated acute myocardial injury in the rat
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.
Activation of NF-kappaB is a critical element in the antiapoptotic effect of anesthetic preconditioning
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.
