Overview

  • Product name

    Phosphatidylcholine Assay Kit (Colorimetric/Fluorometric)
  • Detection method

    Colorimetric/Fluorometric
  • Sample type

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

    Quantitative
  • Sensitivity

    > 0.2 mM
  • Range

    0.2 mM - 2.5 mM
  • Assay time

    0h 40m
  • Product overview

    Phosphatidylcholine Assay Kit (Colorimetric/Fluorometric) (ab83377) is a simple convenient means of measuring phosphatidylcholine in a variety of biological samples. This assay uses an enzyme-coupled reaction to hydrolyze phosphatidylcholine to release choline, which subsequently oxidizes the OxiRed probe in order to generate fluorescence (Ex/Em 535 nm 587 nm) and absorbance (570 nm). This assay measures phosphatidylcholine in the range of 0.1 to 10 nmol per sample.
    Visit our FAQs page for tips and troubleshooting.

  • Notes

    Phosphatidylcholine (PC) is a phospholipid which incorporates choline as the headgroup of the lipid. PC is a major constituent of biological membranes and is involved in cell signaling through release of choline by phospholipase D leaving the second messenger phosphatidic acid.

    PC is present in serum at ~0.2-2.5 mM (~50-200 mg/dL).

  • Platform

    Microplate reader

Properties

Associated products

Images

  • Phosphatidycholine levels measured fluorometrically in mouse tissue lysates (mg of extracted protein; background signal subtracted, mean of duplicates; +/- SD).

  • Phosphatidycholine levels colorimetrically measured  in cell lysates (background signal subtracted, mean of duplicates; +/- SD).

  • Phosphatidycholine levels colorimetrically measured in rat biological fluids (background signal subtracted, mean of duplicates; +/- SD).

  • Examples of Colorimetric and Fluorometric Phosphatidylcholine standard curves using ab83377.

Protocols

References

This product has been referenced in:

  • Promeneur D  et al. Aquaglyceroporin PbAQP is required for efficient progression through the liver stage of Plasmodium infection. Sci Rep 8:655 (2018). Read more (PubMed: 29330527) »
  • Zhao M  et al. Chronic folate deficiency induces glucose and lipid metabolism disorders and subsequent cognitive dysfunction in mice. PLoS One 13:e0202910 (2018). Read more (PubMed: 30153273) »
See all 8 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Answer

Yes, there is a mild detergent in the assay buffer.
Here is the tissue homogenization protocol:
Start with 10-100 mg of the tissue, add 500-1000 μl (or ˜4-6 volumes) of the assay buffer on ice, homogenize using a Dounce homogenizer (10-50 passes) on ice, until efficient lysis is confirmed, by viewing the cells under the microscope. Spin down the sample and collect the supernatant. Use the eluate for your subsequent assays. Appropriate dilutions of the sample must be tested in order ensure the readings will fall within the linear range of the standard curve.

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Answer

Thank you for contacting us regarding the issue which you have been experiencing when using ab83377.

Our development team has reviewed this issue and could not find any specific information about interference from this drug. However they suggest that the following may be occurring. In reviewing the principle behind this assay we see that one of the intermediates in this reaction is choline. It may be that the Hexadecyl phosphocholine might be getting converted into choline resulting in the increased background. It may be valuable to compare samples with lower and higher treatment doses as well as different time points to determine if there is any effect on background resulting from the possible conversion ofHexadecyl phosphocholine to choline.

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 phone call. 
Any treatment on the cells should be performed before the protocol is begun.  Cell lysates can be prepared using the following protocol:

Start with 2 x 106 cells and suspend the cell pellet in 500 uL of assay buffer on ice.

Homogenize using a Dounce homogenizer (10-50 passes) on ice.  Confirm efficient lysis by viewing the cells under a microscope.

Spin down the sample and collect the supernatant. 

Use the eluate in the Phosphatidylcholine Assay.  The sample dilution will need to be optimized.
I hope this helps, please let me know if you need any additional information or assistance.

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Answer

Thank you for your enquiry regarding ab83377. This kit has been tested and characterized for detecting phosphatidylcholine. At this point we do not have such data (reactivity with lysophosphatidylcholine) and this will have to be specifically determined by the end user. If you need any further assistance in the future, please do not hesitate to contact me.

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Answer

Thank you for contacting us. The lab has not tested this kit for use on bile samples, but it may be suitable. The bile samples can be stored at -80C prior to use and diluted in the provided assay buffer. 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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