概述

  • 产品名称
    Free Fatty Acid Assay试剂盒- Quantification
    参阅全部 Fatty acid 试剂盒
  • 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 (FA) in various mammalian and other samples, such as serum, plasma and other body fluids, food, growth media, etc.


    In this assay, FA 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)



    Visit our FAQs page for tips and troubleshooting.


    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.

  • 说明

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

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 40 Publications for this product

客户评价及客户问答

31-34 of 34 Abreviews or Q&A

Answer

I have had a response from the lab about you inquiry concerning the detection bandwidth for the ab6534 kit. Please check if your filter has a bandwidth option. If it does have a +/- 10 bandwidth option you can use this filter and still capture 100% of the signal. However if it doesn't you may be able to capture ~ 75-85% of the signal.

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Answer

As you have detailed the kit measures FFA content in various biological samples (serum, plasma and other body fluids, food, growth media, etc) by enzymatically converting the fatty acids to their CoA derivatives with are oxidised ant their subsequent colour generation measured. As long as there are Fatty Acids within your sample the kit should be able to detect them (detection limit of 2 µM). Please note not the kit is not suitable to detect short chain fatty acids below 8 carbons. The kit is multiple purpose. As long as the components are stored as directed with the protocol. Would advise avoiding frequent freeze-thaw cycles by aliquotting sensitive components into working volumes and storing it at -20°C. This would be best to ensure the activity of the components. Using the kit in this way over 2 weeks should not affect the activity.

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Question
Answer

This kit can be used with tissue samples. The tissue preparation instructions are as follows: 10 mg tissue samples can be extracted by homogenization with 200 µl of chloroform-Triton X-100 (1% Triton X-100 in pure chloroform) in a microhomogenizer. Then spin the extract 5-10 minutes at top speed in a microcentrifuge. Collect organic phase (lower phase), air dry at 50°C to remove chloroform. Vacuum dry 30 min to remove trace chloroform. Dissolve the dried lipids (in Triton X-100) in 200 µl of Fatty Acid Assay Buffer by vortexing extensively for 5 minutes (Note: The solution may be slightly turbid or opalescent, but this does not affect the assay). The extraction procedure can be proportionally scaled up if larger amount of sample is desired. Use 1- 50 µl of the extracted sample per assay.

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Answer

We can confirm that this Kit will not be suitable to use with fatty acids less than 8 carbons or short chain fatty acids. This kit can detect Octanate or long chain fatty irrespective of species of the organism.

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31-34 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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