Creatine kinase (CK) Colorimetric Assay Kit

    • Biochemical Assay Kit-Elabscience
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    • Biochemical Assay Kit-Elabscience
    • Biochemical Assay Kit-Elabscience

      Catalog number:E-BC-K558

      Size:
      • 50 Assays
      • 100 Assays
      Qty:
      - +
      Price: $350

      Detection method: Colorimetric method

      Detection instrument: Spectrophotometry

      Valid period: 3 months

      Lead Time: 7~10 daysWelcome to order from local distributors.

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      General information

      Detection significance

      Creatine kinase (CK), also known as creatine phosphokinase (CPK) or phosphocreatine kinase, is an enzyme (EC 2.7.3.2) expressed by various tissues and cell types. It can transfer phosphate group from phosphocreatine to ADP and catalyze the production of ATP. Creatine kinase plays a key role in cell energy metabolism.

      Detection principle

      Creatine kinase (CK) catalyze creatine phosphate and ADP to produce creatine and ATP. Hexokinase catalyze creatine and glucose to produce glucose-6-phosphate. Glucose-6-phosphate dehydrogenase (G-6-PD) catalyze glucose-6-phosphate and NADP+ to produce NADPH which have a maximum absorption peak at 340 nm. The CK activity can be calculated by measuring the OD values at 340 nm.

      The key point

      1.   Samples should not contain Fe3+ and Cu2+, or it will inhibit the activity of G-6-PD.

      2.   Avoid moderate or severe hemolysis of samples. When the degree of hemolysis is high, red blood cells will release AK, ATP, G-6-PD and so on, which will affect the results.

      3.   The activity of CK is unstable, temperature and light may lead to the loss of enzyme activity. So the sample should be detect as soon as possible after collection, or preserve the samples on ice for before detection.

      4.   When the reagent 1 is taken, it is recommended to avoid the contamination of reagent.

      Operation procedures

      Operation steps

      1)   Blank tube: Take 1000 μL of reagent 1, 50 μL of double distilled water into an EP tube.

      Sample tube: Take 1000 μL of reagent 1, 50 μL of sample into EP tubes.

      2)   Mix fully and incubate at 37℃ for 5 min.

      3)   Add 100 μL of preheated reagent 2.

      4)   Mix fully and incubate at 37℃ for 2 min.

      5)   Set to zero with blank tube and measure the absorbance at 340 nm with 1 cm quartz cuvette at initial absorbance (A1) and 5 min (A2), respectively. Calculate the △A=A1-A2.

      Operation table

       

      Blank tube

      Sample tube

      Reagent 1 (μL)

      1000

      1000

      Double distilled water (μL)

      50

       

      Sample (μL)

       

      50

      Mix fully and incubate at 37 for 5 min.

      Reagent 2 (μL)( preheat in 37 for 10 min)

      100

      100

      Mix fully and incubate at 37 for 2 min. Set to zero with blank tube and measure the absorbance at 340 nm with 1 cm quartz cuvette at initial absorbance (A1) and 5 min (A2), respectively. Calculate the A=A2-A1.

      Performance characteristics

      Technical parameter

      Detection range 3.7-160 U/L Average inter-assay CV 8.1%
      Sensitivity 3.7 U/L Average intra-assay CV 5.2%

      Example analysis

      Detect 10% rat heart tissue homogenate (the concentration of protein is 6.13 gprot/L, dilute for 2 times), 10% rat kidney tissue homogenate (the concentration of protein is 7.84 gprot/L) and 10% rat brain tissue homogenate (the concentration of protein is 4.66 gprot/L, dilute for 2 times) according to the protocol, the result is as follows:

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