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| Product Name | Creatine Kinase (CK) Activity Colorimetric Assay Kit |
| Catalog No. | ETR-HMM-0060 |
| Description | A colorimetric enzymatic assay for quantifying creatine kinase activity in biological samples. CK catalyzes the reversible phosphorylation of creatine using ATP; in the coupled assay, the generated ADP activates the hexokinase-glucose-6-phosphate dehydrogenase enzyme cascade, resulting in NADH production detected at 340 nm. |
| Intended Use | Quantitative CK activity measurement for cardiac injury biomarker research, skeletal muscle disease studies (muscular dystrophy), rhabdomyolysis assessment, and sports medicine research monitoring muscle damage. |
| Principle / Technology | CK catalyzes the reaction: Phosphocreatine + ADP ⇌ Creatine + ATP (reverse reaction used in the IFCC-recommended method); in the coupled assay, CK phosphorylates ADP to ATP using creatine phosphate; the produced ATP is used by hexokinase to phosphorylate glucose to glucose-6-phosphate; glucose-6-phosphate dehydrogenase then oxidizes G6P with concomitant reduction of NADP+ to NADPH, measured kinetically at 340 nm. |
| Detection Method | UV absorbance kinetic measurement at 340 nm using microplate reader with UV capability or conventional spectrophotometer |
| Sample Type | Serum, plasma, cell lysates (cardiac myocytes, skeletal muscle cells, smooth muscle cells, brain tissue), tissue homogenates |
| Performance Range / Specifications | CK activity range: 1–2,000 U/L; typical serum reference range 25–200 U/L (sex-dependent); one unit defined as amount producing 1 μmol NADPH per minute at 37°C; kinetic linearity over 3–5 minutes |
| Sensitivity / LOD | Lower detection limit: approximately 0.5 U/L in serum or 0.1 mU per reaction |
| Specificity | The CK-coupled assay is highly specific for creatine kinase; adenylate kinase interference is eliminated by addition of AMP and diadenosine pentaphosphate (Ap5A) to the assay mixture; CK isoenzymes (CK-MM, CK-MB, CK-BB) are all measured by this total CK method |
| Reaction Conditions / Protocol | Equilibrate all kit components and samples to 37°C; add 10 μL sample to microplate; add 200 μL pre-warmed Working Reagent (all assay components mixed as instructed); mix briefly; incubate at 37°C for 1 minute (activation); begin kinetic absorbance reading at 340 nm every 30 seconds for 3 minutes; calculate ΔOD/min from linear portion of kinetic trace; activity (U/L) = ΔA340/min × total volume / (ε_NADPH × sample volume × light path) |
| Components / Formulation | Reagent 1 (creatine phosphate, HEPES buffer, EDTA, Ap5A, AMP, glucose, magnesium acetate), Reagent 2 (ADP, hexokinase, glucose-6-phosphate dehydrogenase, NADP+, N-acetylcysteine), CK Calibrator (lyophilized human serum-based, traceable reference), reconstitution buffer |
| Storage Conditions | Lyophilized calibrator at -20°C for 12 months; Reagent 1 and Reagent 2 at 2–8°C protected from light for 12 months; working reagent (mixed R1 + R2) stable 4 hours at 37°C |
| Shelf Life | 12 months from date of manufacture |
| Package Specifications | 200 assays, 500 assays, 1,000 assays (96-well/cuvette format) |
| Product Form | Lyophilized calibrators with liquid reagent components |
| Quality Control | Each lot validated against certified CK reference materials from international reference labs; within-run precision CV <2%; recovery 95–105% of spike added CK to serum; Ap5A inhibitor confirmed to suppress adenylate kinase interference >98% |
| Key Features | IFCC-recommended coupled enzymatic method with international traceability; Ap5A adenylate kinase inhibitor eliminates major interference source; liquid reagents reduce preparation time; applicable to both 96-well plate readers and standard cuvette spectrophotometers |
For research use only, not for clinical use.
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