Overview

  • Product name

    L-Carnitine Assay Kit
  • Detection method

    Colorimetric/Fluorometric
  • Sample type

    Urine, Serum, Plasma, Other biological fluids, Cell Lysate, Tissue Lysate
  • Assay type

    Quantitative
  • Range

    1 µM - 200 µM
  • Assay time

    0h 30m
  • Species reactivity

    Reacts with: Other species, Mammals
  • Product overview

    Abcam's L-Carnitine Assay Kit is a simple convenient means of measuring free L-Carnitine in biological samples such as serum. The assay transfers an acetyl group from CoA to carnitine and the free CoA formed is further processed with subsequent oxidation of the OxiRed probe to give fluorescence (Ex/Em 535 nm 587 nm) and absorbance (570 nm). The normal range for serum L-Carnitine is ~20-100 µM. The detection sensitivity is ~1 µM for the fluorometric assay and ~10 µM for the colorimetric assay.
    Visit our FAQs page for tips and troubleshooting.

  • Notes

    Carnitine is a quaternary ammonium compound biosynthesized from the amino acids lysine and methionine. It is required for transport of fatty acids into the mitochondrial matrix via the carnitine/acylcarnitine shuttle where beta-oxidation occurs, acetate is generated and the acetate utilized in the TCA cycle for the generation of energy. L-Carnitine is often sold as a nutritional supplement. Carnitine exists in two stereoisomers. Only L-carnitine is biologically active.

  • Platform

    Microplate reader

Properties

Associated products

Images

  • Fluorimetric standard curve: mean of duplicates (+/- SD) with background readings subtracted.

  • L-Carnitine measured in biologicals showing concentration (micromolar).

  • Example of L-Carnitine standard curve obtained using ab83392.

Protocols

References

This product has been referenced in:

  • Yaligar J  et al. Longitudinal metabolic imaging of hepatocellular carcinoma in transgenic mouse models identifies acylcarnitine as a potential biomarker for early detection. Sci Rep 6:20299 (2016). Read more (PubMed: 26831370) »
See 1 Publication for this product

Customer reviews and Q&As

1-10 of 17 Abreviews or Q&A

Question
Answer



This kit will be very suitable for use with EDTA plasma samples.

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Abreviews
L-carnitine concentration was measured using fluorimetry in a yeast strain capable of carnitine biosynthesis, both with and without supplementation with the carnitine precursor gamma-butyrobetaine (gBB).
The yeast were lysed using glass beads in assay buffer. The background was higher than in the fluorimetric standard curve, likely due to interfering metabolites, possibly choline. The increased carnitine detection in gBB-supplemented cells suggests that kit ab83392 can be used for comparative carnitine measurements in cells other than human origin, including in yeasts however, the high background levels complicate absolute quantification.

Abcam user community

Verified customer

Submitted Jun 03 2016

Answer

The ab138889 CoA detection kit assay buffer is 100 mM Na phosphate buffer, pH = 6. The RIPA buffer may be an issue with the other two kits, since they are enzyme-based, and that kind of assay is generally incompatible with detergents. Your best approach will be to dilute the samples as much as you can with the provided buffers, while maintaining the diluted sample OD values (for the carnitine assay) and RFU values (for the other two) remain within the range of the standards. So, you will need to test serial dilutions of, for instance, 1/2, 1/4, and 1/8.

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Answer

The instructions in the protocol are correct, and the values are too, 0, 2, 4, 6, 8, and 10 nmol per well. Here is the calculation:

Starting with the 1 mM standard carnitine solution, calculate the moles per microliter.

1 millimole per liter = 1 nanomole (nmol) per microliter.

Add the volumes indicated in the protocol per well, 0, 2, 4, 6, 8, 10 ul, to get the nanomoles per well listed in the protocol.

I hope that makes sense. There is sometimes confusion between nanomoles (amount, nmol) and nanomolarity (concentration, nM). The concentration can be calculated from the amount and the volume, for example, 10 nanomoles per 50 ul equals 200,000 nanomoles (10 x 20,000) per 1,000,000 microliters (50ul x 20,000) = 0.2 millimoles per liter = 0.2 mM (concentration).

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Answer

Thank you for contacting us.

We are not ISO registered company which is why we do not have CE certification. We can however provide certificate of analysis, if requested. I don't think CE certification is required for research products. Our products conform to European law which is why we sell it throughout Europe without any problem.

This kit will work for assaying L-Carnitine irrespective of species of 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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Answer

Thank you for your message and for your patience.

I have already received information to confirm that this kit has been optimized to measure free L-carnitine and so regrettably we cannot guarantee or advice regarding measurement of total L carnitine.

I had been waiting to receive a more precise list of on mammalian species in which this has been tested but I am sorry I have not yet received a response. I will let you know as soon as I receive more information.

If you have any further questions, please do not hesitate to contact me.

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Answer

Thank you for your enquiry regarding ab83392.

My colleague Kate has been out of the office this morning. Here are some links which show these levels:

http://www.kidney.org/professionals/KDOQI/guidelines_ped_ckd/cpr9.htm

http://www.sciencedirect.com/science/article/pii/S000991200100234X

http://humrep.oxfordjournals.org/content/23/7/1602.full

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

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Answer

Thank you for your inquiry.

This kit should be very specific for L-Carnitine since the enzymes used are specific to it. Also the reaction mimics the biological activity of L-Carnitine and only L-Carnitine is active not D-Carnitine.

It has not been experimentally tested if this kit also detects D-Carnitine, but if it does is should be only in negligible amount.

I am sorry I could not provide a straight forward answer on this occasion and hope this information is nevertheless helpful.

I wish you good luck with your research and a good weekend.

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Answer

Thank you for contacting Abcam

Upon arrival the kit has to be stored at -20C. Certain key components in the kit are lyophilized so the kit may be stable. However I cannot guarantee this and you will have to test whether it is still working.

The carnitine assay buffer should be in the kit that you received (you should have received 1x25ml of it) and you can use any standard 96 well plate to perform the assay in.

Please let me know if there is anything else I can help you with.

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Answer

Thank you for contacting Abcam regarding ab83392. Unfortunately, the specific details of the reaction mechanism and enzyme names are proprietary information in this case. I can only provide the following: Acetyl CoA + Carnitine ---> Free CoA + Acetylated Carnitine. Free CoA + Oxired probe ---> Proportionate increase in color or fluorescence. I apologize that I am not able to provide more specific details. I hope that this information is helpful. Please do not hesitate to contact us if you have any additional questions.

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1-10 of 17 Abreviews or Q&A

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