Creatine Kinase Activity Assay Kit (Colorimetric) (ab155901)

Reviews (2)Q&A (8)References (14)
Functional studies - Creatine Kinase Activity Assay Kit (ab155901)
  • Functional Studies - Creatine Kinase Activity Assay Kit (Colorimetric) (ab155901)
  • Functional studies - Creatine Kinase Activity Assay Kit
  • Figure 1

Key features and details

  • Assay type: Enzyme activity
  • Detection method: Colorimetric
  • Platform: Microplate reader
  • Assay time: 1 hr
  • Sample type: Adherent cells, Plasma, Serum, Suspension cells, Tissue, Tissue Homogenate
  • Sensitivity: 1 mU/well

Overview

  • Product name

    Creatine Kinase Activity Assay Kit (Colorimetric)

  • Detection method

    Colorimetric

  • Sample type

    Serum, Plasma, Tissue, Adherent cells, Suspension cells, Tissue Homogenate

  • Assay type

    Enzyme activity

  • Sensitivity

    < 1 mU/well

  • Range

    2 nmol/well - 10 nmol/well

  • Assay time

    1h 0m

  • Species reactivity

    Reacts with: Mammals, Other species

  • Product overview

    Creatine Kinase Activity Assay Kit (Colorimetric) ab155901 uses a high-throughput adaptable, simple and sensitive assay for creatine kinase (CK) activity.


    In the creatine kinase assay protocol, creatine kinase (CK) converts creatine into phosphocreatine and ADP. The phosphocreatine and ADP then react with the CK enzyme mix to form an intermediate, which reduces a colorless probe to a colored product with strong absorbance at λ= 450 nm.


    This assay kit can detect CK activity to less than 1 mU.


    Creatine kinase assay protocol summary:
    - add samples and standards to wells
    - add reaction mix 
    - analyze every 1-2 min for 10-40 min with microplate reader in kinetic mode at 37ºC

  • Notes

    Creatine Kinase (CK) also known as creatine phosphokinase (CPK) and ATP: creatine N-phosphotransferase is a common cellular enzyme (EC 2.7.3.2). It catalyzes the reversible conversion of creatine and ATP into ADP and phosphocreatine.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components Identifier 100 tests
    ATP Orange 1 vial
    CK Assay Buffer WM 1 x 25ml
    CK Developer Red 1 vial
    CK Enzyme Mix Green 1 vial
    CK Positive Control Purple 1 vial
    CK Substrate Blue 1 x 1ml
    NADH Standard Yellow 1 vial

  • Research areas

Associated products

    Images

    • Functional studies - Creatine Kinase Activity Assay Kit (ab155901)
      Functional studies - Creatine Kinase Activity Assay Kit (ab155901)

      Creatine Kinase Activity measured in various samples.  

      Initial protein concentration for tissue lysates varied from 1 mg/mL to 3 mg/mL. 40 μl of filtered tissue lysates were used. 30 μl of filtered biological fluids were used.

    • Functional Studies - Creatine Kinase Activity Assay Kit (Colorimetric) (ab155901)
      Functional Studies - Creatine Kinase Activity Assay Kit (Colorimetric) (ab155901)Sengle, Gerhard et al., PLoS genetics?vol. 11,6 e1005340., Table 1, doi:10.1371/journal.pgen.1005340
      Creatine kinase activities in blood samples from wildtype and Fbn2 null mice.
      Mean +/- standard deviation for a given genotype and age is shown, except where only one animal was tested. N = number of animals tested. Units are nmol NADH/min/ml.
    • Functional studies - Creatine Kinase Activity Assay Kit
      Functional studies - Creatine Kinase Activity Assay Kit

      Typical NADH standard calibration curve using colorimetric reading.

    • Figure 1
      Figure 1

      Creatine kinase activity tested in 5 µl human serum and 192 ng rat heart lysate. Assay performed following kit protocol.

    References (14)

    Publishing research using ab155901? Please let us know so that we can cite the reference in this datasheet.

    ab155901 has been referenced in 14 publications.

    • McRae NL  et al. Genetic reduction of the extracellular matrix protein versican attenuates inflammatory cell infiltration and improves contractile function in dystrophic mdx diaphragm muscles. Sci Rep 10:11080 (2020). PubMed: 32632164
    • Ganeshan K  et al. Energetic Trade-Offs and Hypometabolic States Promote Disease Tolerance. Cell 177:399-413.e12 (2019). PubMed: 30853215
    • Song TH  et al. Dendrobine targeting JNK stress signaling to sensitize chemotoxicity of cisplatin against non-small cell lung cancer cells in vitro and in vivo. Phytomedicine 53:18-27 (2019). PubMed: 30668397
    • Zhang Z  et al. Astragalus mongholicus (Fisch.) Bge Improves Peripheral Treg Cell Immunity Imbalance in the Children With Viral Myocarditis by Reducing the Levels of miR-146b and miR-155. Front Pediatr 6:139 (2018). PubMed: 29977885
    • Luptak I  et al. Decreased ATP production and myocardial contractile reserve in metabolic heart disease. J Mol Cell Cardiol 116:106-114 (2018). PubMed: 29409987
    View all Publications for this product

    Customer reviews and Q&As

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    Creatine Kinase Activity Assay Kit (Colorimetric)

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    Abcam user community

    Verified customer

    Submitted Jan 24 2018

    Creatine kinase activity Assay with culture media

     Average Good 4/5 (Ease of Use)
    Abreviews
    abreview image
    I used phenol red free DMEM media with 1% ITSS and 1% antibiotic/antimycotic. Dosed media was incubated with sheep cardiomyocytes for 48 hours and collected. Media was diluted 1:2 with molecular grade water and added to the kits reaction mix. Absorption readings were taken every 2 minutes for 40 minutes.


    Abcam user community

    Verified customer

    Submitted Sep 08 2017

    Question



    Inquiry: I was wondering if the Creatine Kinase Activity Assay Kit could be used with blood collected from tail clips from mice? According to the protocol, the amount of whole blood needed for the assay is 10uL, correct? Thank you

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    Answer

    Thank you for contacting us.





    The sample volume window is 1-50 ul. How much blood is used depends on the CK activity in the sample. When we used human serum (see data fig 1b on our datasheet), 5ul worked well.



    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.



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    Question

    The ab155901 Creatine Kinase activity kit is listed to work with "Serum, Plasma, Tissue, Adherent cells, Suspension cells, Tissue Homogenate", however, the protocol booklet only appears to list instructions for tissues or cells, for example, section B.2. Sample Preparation.​ If I am to use plasma, would I start section B.2. simple by adding "1-50 ul sample per well"?

    Many Thanks,


    Read More
    Answer

    Thank you for your enquiry.
    I can confirm that when using the ab155901 Creatine Kinase Activity Assay Kit there is no special sample preparation required for plasma samples.
    The sample volume used per well will need to be optimized to get values within the linear range of the standard curve.
    I hope this will be helpful.

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    Question

    I want to use this in a cell culture supernatant, are there any reagents in media which would be incompatible with this kit?

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    Answer

    Thank you for contacting us.







    Creatine kinase in cells is either cytosolic or mitochondrial. The enzyme is not released into the medium under normal conditions. SO we would expect to see very low or no activity in the medium under normal healthy culture conditions. The medium as such should not interfere in the assay, althouugh serum in the medium might contain CK enzyme in which case there can be interference from this.


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    Question

    I bought your Creatine kinase kit (ab155901) and the data sheet says that I need to homogenise my cells with 100UL of provided assay buffer. My question is , DO I need to add anti-phosphates and anti-proteases to that assay buffer? Can I keep this assay buffer in the fridge? And what is the chemical composition of that buffer?

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    Answer

    I have contacted the lab and you do not need to add protease of phosphatase inhibitors for our kits unless otherwise stated. This is because, many of these inhibitors could chelate ions or adversely affect the enzymes in the assays.

    Unfortunately, the buffer composition is proprietary and hence cannot be shared.

    Please store the assay buffer as stated in the protocol booklet so at -20ºC. We have not tried storing it at 4ºC and so cannot guarantee that it will work if stored differently to what we recommend.

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    Question

    I have read in the protocol that the tissue should be homogeneized after collection, however, as tissue collection can take some time I would like to know what is the best way to preserve the tissue in the meantime so that enzyme activity is not lost (on ice???).
    Is it possible to freeze the samples and for how long? Is it better to freeze the whole tissue and then homogeneise after thawing or is it better to homogeneise and get the supernantant first and then freeze the supernatant until samples are processed? Is it better to freeze at -20 or -80 degrees?

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    Answer



    We recommend collecting the skeletal muscle samples and quick freeze in liquid nitrogen and store at -80C until they are ready to run all the samples together.

    The tissue samples should be homogenized when they are ready to do the experiment and then a lysate can be prepared freshly.

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    Question

    Positive control and sample absorbances are outside the range of the standards.

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    Answer

    The positive control activity may vary from batch to batch. It is included in the assay to confirm that the assay reagents are performing as expected. If the OD had been 0, then that would be a problem. An OD higher than the highest standard is not. To bring your sample OD values and the positive control OD value within the range of the standards, you will need to dilute them. Testing two or three dilutions for each will be best, to ensure that at least one of them is within the range of the standards.

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    Question

    I’d like to clarify whether the Creatine Kinase Activity Assay Kit (Colorimetric) ab155901 is reactive to Rat CK in plasma samples.

    With kind regards

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    Answer

    The kit is guaranteed to be used with Rat samples. It is tested with rat heart lysates and we predict it will work with plasma samples as well.

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    Question

    Does this kit detect U-type creatine kinase (mitochondrial)? Do you have any data for use with brain lysates?

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    Answer

    This kit will detect total creatine kinase, including U-type. Unfortunately we do not have any specific data produced by the kit with brain homogenates.

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