Overview

  • Product name

    Creatine Assay Kit (Colorimetric/Fluorometric)
  • Detection method

    Colorimetric/Fluorometric
  • Sample type

    Cell culture supernatant, Urine, Serum, Plasma, Other biological fluids
  • Assay type

    Quantitative
  • Sensitivity

    > 0.001 mM
  • Range

    0.001 mM - 10 mM
  • Assay time

    1h 00m
  • Species reactivity

    Reacts with: Mammals, Other species
  • Product overview

    Creatine Assay Kit (Colorimetric/Fluorometric) (ab65339) provides an accurate, convenient measure of creatine in a variety of biological samples.


    In the creatine assay protocol, creatine is enzymatically converted to sarcosine which is then specifically oxidized to generate a product that converts a colorless probe to an intensely red color (λmax = 570 nm), and highly fluorescent (Ex/Em = 538/587 nm) product. Creatine is therefore easily detected by either colorimetric or fluorometric methods. Detection range 0.001–10 mM Creatine.


    Creatine assay protocol summary:
    - add reaction mix to sample and standard wells
    - incubate for 60 min
    - analyze with a microplate reader

  • Notes

    Creatine is an endogenous compound whose function is to maintain a high ATP/ADP ratio, by way of its phosphorylated form and creatine kinase. Creatine supplementation has been used in the treatment of muscular, neurological and neurodegenerative diseases, as well as a sport performance enhancer. Detection of creatine level has wide applications in research and development.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components Identifier 100 tests
    Creatinase (Lyophilized) Blue 1 vial
    Creatine Assay Buffer WM, 1 x 25ml
    Creatine Enzyme Mix (Lyophilized) Green 1 vial
    Creatine Probe (in DMSO) Red 1 x 200µl
    Creatine Standard (10 µmol) (Lyophilized) Yellow 1 vial
  • Research areas

  • Relevance

    Creatine is an endogenous amino acid that occurs in vertebrate tissues and in urine. In muscle tissue, creatine is generally found as its phosphorylated form phosphocreatine. Creatine functions as part of the cell's energy shuttle: the high energy phosphate group of ATP is transferred to creatinine to form phosphocreatinine in the following reaction: Cr + ATP ? P-Cr + ADP. This reaction is reversibly catalized by creatine kinase. In the human body, creatine is synthesized mainly in the liver from arginine, glycine and methionine. Creatine is excreted as creatinine in the urine.

Associated products

Images

  • Creatine measured in cell lysates showing quantity (nmol) per 106 cells

  • Creatine measured in biologicals showing concentration (micromolar)

Protocols

References

This product has been referenced in:

  • Wan ES  et al. Metabolomic profiling in a Hedgehog Interacting Protein (Hhip) murine model of chronic obstructive pulmonary disease. Sci Rep 7:2504 (2017). Read more (PubMed: 28566717) »
See 1 Publication for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Answer

It is correct that this kit can detect phosphorylated and non-phosphorylated creatine.

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Answer

Sorry for any confusion here.
In the assay, creatine is enzymatically converted to sarcosine which is catalyzed by creatinase. This enzyme works at the first step of the detection reaction in the kit. This conversion event is not dependent on phosphorylation of creatine. So in summary this kit can't be used to distinguish between phosphocreatine unphosphorylated creatine.

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Answer

Thank you for your enquiry and interest in our products.

Generally, most plate readers have flexibility in their bandwidth of detection. This is generally in terms of ±10, ±20 and so on. If your machine has the flexibility of at least ±20, you will be able to use your wavelengths instead of the recommended ones.

If you need any further assistance in the future, please do not hesitate to contact me.

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Answer

Perdona por la espera.

He vuelto a comentar el caso con mis compañeros. Me temo que no tienen ninguna sugerencia que hacer respecto a un control positivo de Creatina para usar con el kit. Podríais utilizar Creatina pura en distintas concentraciones, o quizás filtrar las muestras con uno de los filtros de 10KDa recomendados, para optimizar la preparación de la muestra.

Siento no poder ser de más ayuda en esta ocasión. Si tenéis cualquier otra consulta, no dudes en contactarnos de nuevo.

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Answer

Thank you for contacting us.

I have contacted the lab on your behalf. This product will work very well with mouse samples.

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
Answer

Thank you for your phone call.

I can confirm that this kit will not be able to distinguish between phospho-creatine and creatine molecules. The enzyme used in this kit Creatine Amidinohydrolase which converts creatine into Sarcosine.

If you are interested in determining the level of phpspho-creatine then one way is converting all the phospho-creatine to creatine using Creatinine Amidohydrolase enzyme.

By subtracting the value of creatine from total creatine, will give you the phospho-creatine level.

I hope this information will be helpful. Should you have any other question please do not hesitate to contact me.

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Answer

Thank you for contacting us.
I have to confirm that we have not specifically tested with this sample type (tissue) and it is therefore not covered by our guarantee.
This kit should be compatible with all mammalian samples and therefore should also theoretically work with mouse heart cell lysates.
I suggest to homogenize the tissue in assay buffer with a dounce homogenizer. This will have to be optimized by the end user.
High concentrations of proteins in lysates may interfere with the assay. Samples containing proteins may be filtered through a 10k MW cut-off filter:
ab93349, https://www.abcam.com/10kD-Spin-Column-ab93349.html
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
Answer

Thank you for contacting us.

The lab let me know the following:

The assay buffer composition is proprietary information. However, we recommend to use the buffer which comes with each kit, instead of substituting it with another kit’s buffer.

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

Use our products? Submit an Abreview. Earn rewards!
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Answer

Thank you for your inquiry.


The ab65339 kit measures creatine, while the ab65340 kit measures creatinine.


https://www.abcam.com/Creatine-Assay-Kit-ab65339.html


https://www.abcam.com/Creatinine-Assay-Kit-ab65340.html


The Creatine Assay Kit provides an accurate, convenient measure of creatine in a variety of biological samples. In the assay, creatine is enzymatically converted to sarcosine which is then specifically oxidized to generate a product that converts a colorless probe to an intensely red color (detected by spectrophotometry at OD = 570 nm), and highly fluorescent (Ex/Em = 538/587 nm) product. Creatine is therefore easily detected by either colorimetric or fluorometric methods. Detection range 0.001 – 10 mM Creatine.


The Creatinine Assay Kit provides an accurate, convenient measure of creatinine concentration in biological fluids such as serum, urine or CSF. In the assay, creatinine is converted to creatine by creatininase, creatine is converted to sarcosine, which is specifically oxidized to produce a product which reacts with a probe to generate red color (detected by spectrophotometry at OD = 570 nm) and fluorescence (Ex/Em = 538/587 nm). This kit is suitable for serum/plasma creatinine determinations, as well as for urine and other biological samples.


Essentially the only difference is that the creatinine kit involves an additional step involving creatininase to create creatine.


I hope this information helps. Please contact us with any other questions.

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Answer

Thank you for contacting us.

This product has an expiration date of 6 months from date of arrival if stored at the recommended stated on the datasheet (-20C).

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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1-10 of 12 Abreviews or Q&A

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