概述

  • 产品名称
    Free Fatty Acid Assay试剂盒- Quantification
  • 检测方法
    Colorimetric/Fluorometric
  • 样品类型
    Cell culture supernatant, Urine, Serum, Plasma, Other biological fluids, Tissue Extracts
  • 检测类型
    Quantitative
  • 灵敏度
    > 2 µM
  • 检测时间
    0h 40m
  • 产品概述

    Free Fatty Acid Quantification Assay Kit ab65341 provides a convenient, sensitive enzyme-based method for detecting long-chain free fatty acids in various mammalian and other samples, such as serum, plasma and other body fluids, food, growth media, etc.


    In the free fatty acid assay protocol, fatty acids are converted to their CoA derivatives (coenzyme A), which are subsequently oxidized, leading to formation of color/ fluorescence. Fatty acids can then be easily quantified by either colorimetric (spectrophotometry at λ= 570 nm) or fluorometric (at Ex/Em= 535/587 nm).

  • 说明

    This assay detects formation of C-8 (octanoate) and longer fatty acids.

    Review our Metabolism Assay Guide to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.

  • 平台
    Microplate reader

性能

图片

  • Plasma FFA levels was measured using ab65341 in male and female wild-type (WT) or AT2KO mice either on normal diet (ND) or high fat diet (HFD).

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

  • Free Fatty Acid measured in biologicals showing concentration (µM)

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

实验方案

文献

This product has been referenced in:
  • Harm S  et al. Removal of stabilizers from human serum albumin by adsorbents and dialysis used in blood purification. PLoS One 13:e0191741 (2018). Human . Read more (PubMed: 29364955) »
  • Shimazu T  et al. Role of METTL20 in regulating ß-oxidation and heat production in mice under fasting or ketogenic conditions. Sci Rep 8:1179 (2018). Read more (PubMed: 29352221) »
See all 47 Publications for this product

客户评价及客户问答

11-20 of 34 Abreviews or Q&A

Question
Answer

This kit will only detect the unbound free fatty acids in you sample and not any albumin bound free fatty acids,

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Answer


The Free Fatty Acid Quantification Kit (ab65341) allows for the quantification of long-chain free fatty acids in various biological samples, such as serum, plasma and other body fluids, food, growth media. It is able to measure chain lengths of C8 or longer. The kit provides a total fatty acid measurement and would not allow for the measurement of individual different fatty acids.

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Answer

Unfortunately, the components of the buffer in question are considered proprietary. However, the lab has asked that if you have a specific question to ask about the buffer, we will try to do our best to address the problem.

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Answer

Yes, ab65341 has flexible uses so it can be used for both colorimetric and fluorometric, giving the user the choice and convenience of being able to perform and analyze both output types.

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Answer

The “ACS” stands for Acyl-CoA synthesis, as in ACS reagent would be Acyl-CoA synthesis reagent.

I regret to inform you that I am not allowed to disclose the composition of this reagent due to proprietary restrictions.

Fatty Acid (FA) shorter than C8 do not act as good substrates for the CoA conversion. Therefore, you could say that beginning step 3, all steps are crucial.

CoA derivatives are oxidized by the enzymes present in the enzyme mix which leads to the generation of color or signals.

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Answer

The assay buffer used in ab65341 contains NP-40.

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Answer



Yes, these plates should be fine for your assay.

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Answer



1.) Since the fluorometric assay is more sensitive than the colorimetric one, your standards might just plateau off if a higher concentration for the standard curve is used. I would say that 1 nmol/well of the standard as mentioned in the datasheet is the maximum that should be used for a fluorometric assay.

If the samples are expected to contain a high amount of free fatty acids, I recommend to dilute them in order to get results that lay within the standard curve.

2.) Yes, by all means use oleic acid standards in parallel with the PA standards that are provided. Basically the results should be the same with either, since this kit is suitable for all C-8 (octanoate) and longer fatty acids.

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Answer

Yes, this kit can be used for all C-8 (octanoate) and longer fatty acids assay. Since oleic acid fits into this category, you can use this assay kit. But please leer in mind that any other long chain fatty acids also present in the cells would be assayed for by the kit. There would be no way to differentiate one from the other.

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Answer

As some of the components are quite sensitive like Fatty Acid enzyme mix, I am afraid that the kit will rather not work. However, they may be lucky: they could only perform an ELISA with the standard to check if the components are still working.

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11-20 of 34 Abreviews or Q&A

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

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