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Cholesterol/Cholesteryl Ester Quantitation Kit (ab65359)

Q&A (29)Specific References (4)

Overview

  • Product nameCholesterol/Cholesteryl Ester Quantitation KitSee all Cholesterol/Cholesteryl Ester kits ...
  • Tests
    100 x 1 assay
  • Sample type
    Cell culture supernatant, Urine, Serum, Plasma, Other biological fluids, Tissue Extracts
  • Assay typeQuantitative
  • Assay time
    1h 00m
  • Product overview

    Abcam's Cholesterol/Cholesteryl Ester Quantitation Kit provides a simple method for sensitive quantification of free cholesterol, cholesteryl esters, or both by colorimetric or fluorometric methods. Majority of the cholesterol in blood is in the form of cholesteryl esters which can be hydrolyzed to cholesterol by cholesterol esterase. Cholesterol is then oxidized by cholesterol oxidase to yield H2O2 which reacts with a sensitive cholesterol probe to produce color (λmax = 570 nm) and fluorescence (Ex/Em = 535/587 nm). The assay detects total cholesterol (cholesterol and cholesteryl esters) in the presence of cholesterol esterase or free cholesterol in the absence of cholesterol esterase in the reaction. Cholesteryl ester can be determined by subtracting the value of free cholesterol from the total (cholesterol plus cholesteryl esters).
    Visit our FAQs page for tips and troubleshooting.

  • Tested applicationsFunctional Studies more details

Properties

  • RelevanceCholesterol is an essential molecule in all animal life. It is involved in both normal development and diseases. A high level of serum cholesterol is an indicator for diseases such as heart disease. Cholesteryl ester is an ester of cholesterol. The ester bond is formed between the carboxylate group of a fatty acid and the hydroxyl group of cholesterol.

    • Applications

    Our Abpromise guarantee covers the use of ab65359 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    Application Notes
    Functional Studies FuncS

    Cholesterol/Cholesteryl Ester Quantitation Kit images

    • Functional Studies - Cholesterol/Cholesteryl Ester Quantitation Kit (ab65359)
      Functional Studies - Cholesterol/Cholesteryl Ester Quantitation Kit (ab65359)
      Detection of Cholesterol/Cholesteryl Ester Using Cholesterol Quantitation Kit (ab65359). Cholesterol/Cholesteryl Ester was quantified using the kit by colorimetric (A) and fluorometric (B) methods according to the kit instructions. Background from the control reaction (without cholesterol) has been subtracted from each value. Note: Fluorometric assay is over 10 fold more sensitive than colorimetric assay.

    Protocols

    References for Cholesterol/Cholesteryl Ester Quantitation Kit (ab65359)

    This product has been referenced in:
    • Shao W  et al. Curcumin prevents high fat diet induced insulin resistance and obesity via attenuating lipogenesis in liver and inflammatory pathway in adipocytes. PLoS One 7:e28784 (2012). Functional Studies . Read more (PubMed: 22253696) »
    • Malur A  et al. Rituximab therapy in pulmonary alveolar proteinosis improves alveolar macrophage lipid homeostasis. Respir Res 13:46 (2012). Read more (PubMed: 22697800) »

    See all 4 Publications for this product

    Publishing research using ab65359 ? Please let us know so that we can cite the reference in this datasheet

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    Submit an Abreview for ab65359 Submit a Question for ab65359

    Displaying 1 - 10 of 29 results for Abreviews and Q&A

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    Votes: Highest First

    Will using trypsin to collect and harvest cells off of plates interfere with the assay?



    Trypsin is a protease so it shouldn't have any effect on the cholesterol in the sample.


    Inquiry: Dear Product Manager, I would like to know if your cholesterol/cholesteryl ester assay kit works for analysis ergosterol/ergosteryl ester. Did you test that? Qian

    Thank you for contacting us.

    We have not tested the kit this way, and are not sure whether it would work for ergosterol/ergosterol ester. Please do not hesitate to contact us if you need any more advice or information.

    Hi,
    More question on the kit.
    When I use it to measure OD at 570nm, I saw some cloud in solution ( your instruction mention it too), I checked OD at 570 after add sample, before add probe, and OD was about 0.4. I think we need to consider this bac...

    Read More

    Thanks for your inquiry. The datasheet in step 2, calls for dissolving dried lipids in 200 µl of Cholesterol Assay Buffer by sonicating or vortexing until homogeneous. At this stage, it is OK if the solution becomes cloudy. After extraction, one mus...

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    Is there something I can add to stop the reaction? From the time I start measuring the with the fluorescent detection to the time I take it out, the color has changed so I don't think measurement is reliable.

    Thank you for contacting us. I forwarded your inquiry to the lab and got the reply below:

    This is an end point assay that will not require a stop reagent. The client will incubate his samples for 60 min and then take the readings. He is placing t...

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    I have a question regarding the application of buffers. I’m using the cholesterol/chol. ester quantification Kit ab65359 and I am interested in free fatty acid quantification Kit ab65341. My question is focussing on the extraction of lipids and the ...

    Read More

    Thank you for contacting us.

    Unfortunately, the assay buffers in both kits have different components and pH. Therefore they cannot be used interchangeably.

    I hope this information is helpful to you. Please do not hesitate to contact us i...

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    Dear sir,

    We are using HDL and LDL/VLDL Cholesterol Assay Kit ab65390

    Thanks

    Thank you for confirming what Kit is used for your experiments.

    I contacted the laboratory to find the right answer for your question.

    Regrettably we cannot give you a guaranteed answer. We believe the chylomicrons would be with the LDL/...

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    Dear Sir,

    Where would chylomicron be after precipitation (in supernatant or precipitate)? Usually we don’t ask our student to fast before giving blood

    Best greetings

    Thank you for contacting us.

    Could you confirm if you want to use the Cholesterol/Cholesteryl Ester Quantitation Kit (ab65359)?

    This kit is guaranteed to work with serum and plasma samples.

    Serum is the liquid fraction of whole ...

    Read More

    Dear Tech,

    According to the protocol for this cholesterol/cholesteryl ester quantitation kit, chloroform : isopropanol: NP-40 (7:11:0.1) is recommended to use for extraction with tissue sample. I would like to ask whether the sample is suitable f...

    Read More

    Thank you for contacting us.

    The protocol for ab65359 is optimized with ChCl3, isopropanol and NP-40 which are important elements in tissue sample extraction. The 3:1 mixture of Chloroform:methanol might work, however, it is hard to tell the effi...

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    Thank you for looking into this issue. I have received a response from the customer regarding the questions raised in your email, please see below:

    I used 0.4ul per sample for the probe.

    Ignore the black readings. On the original rea...

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    Thank you for your reply with further information.

    I have added a FOCR kit to one of your next orders so it should be with your shortly.


    I hope the new kit works fine for them. Just let me know if they have further questions.

    Problems with the standard curve

    Apologies for the delay in getting back to you.

    - Some of the data looks supersaturated, could you confirm what amount of the probe was used. Use of a high amount of probe generates more background in fluorescent assays.

    - What do th...

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"