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    products/assay-kits/free-fatty-acid-assay-kit-quantification-ab65341.pdf

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Cardiovascular Lipids / Lipoproteins Fatty Acids Synthesis
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Free Fatty Acid Assay Kit - Quantification (ab65341)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (1)Q&A (33)References (242)

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Functional Studies - Free Fatty Acid Assay Kit - Quantification (ab65341)
  • Functional studies - ab65341
  • Func S-1
  • ab65341 Free Fatty Acid Quantification Kit
  • ab65341 Free Fatty Acid Quantification Kit

Key features and details

  • Assay type: Quantitative
  • Detection method: Colorimetric/Fluorometric
  • Platform: Microplate reader
  • Assay time: 60 min
  • Sample type: Cell culture supernatant, Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine
  • Sensitivity: 2 µM

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Overview

  • Product name

    Free Fatty Acid Assay Kit - Quantification
    See all Free Fatty Acid kits
  • Detection method

    Colorimetric/Fluorometric
  • Sample type

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

    Quantitative
  • Sensitivity

    > 2 µM
  • Assay time

    0h 60m
  • Product overview

    Free Fatty Acid Assay Kit ab65341 uses 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 (λ= 570 nm) or fluorometric (Ex/Em= 535/587 nm) methods. Palmitic acid is used to generate a standard curve.


    Free fatty acid assay protocol summary:
    - add samples and standards to wells
    - add reaction mix and incubate for 30 min at 37ºC
    - analyze with microplate reader


    Δ Note: Assay time does not include preparation of the samples.


    Chinese protocol available. See protocols section below.

  • Notes

    This product is manufactured by BioVision, an Abcam company and was previously called K612 Free Fatty Acid Quantification Colorimetric/Fluorometric Kit. K612-100 is the same size as the 100 test size of ab65341.

    This assay detects 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.
     

    How other researchers have used Free Fatty Acid Assay Kit ab65341

    This Free Fatty Acid assay kit has been used in publications in a variety of sample types, including:
    - Human: cell culture lysates1, serum2
    - Mouse: plasma and liver tissue3, serum4, plasma5, heart tissue6, liver7
    - Rat: serum8, plasma9, cytosol of articular chondrocyte primary cell cultures10
    - Cell culture medium11 
    - C reinhardtii algae12
    - Drosophila13
    - C elegans14

    References: 1 - Ali A et al 2018, Phokrai P et al 2018; 2 - Tahapary DL et al 2018; 3 - Becares et al 2019; 4 - Pan J et al 2019, Maatta J et al 2018, Rohm M et al 2018, Shimazu T et al 2018; 5 - Woo M et al 2018, Tian X et al 2017; 6 - Liu W et al 2017; 7 - Hao et al 2017, Rui W et al 2016; 8 - Honore SM et al 2018; 9 - Van der Werf et al 2018;10 - Lee SW et al 2018; 11 - Kostrzewski T et al 2017; 12 - Ramanan R et al 2018; 13 - Scopelliti A et al 2019; 14 - Pollard AK et al 2019

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components Identifier 100 tests 2000 tests
    ACS Reagent Blue 1 vial 20 vials
    Enhancer Purple 1 x 200µl 20 x 200µl
    Fatty Acid Assay Buffer WM 1 x 25ml 20 x 25ml
    Fatty Acid enzyme mix Green 1 vial 20 vials
    Fatty Acid Probe (in DMSO) Red 1 x 200µl 20 x 200µl
    Palmitic Acid Standard (1nmol/µl) Yellow 1 x 300µl 20 x 300µl
  • Research areas

    • Cardiovascular
    • Lipids / Lipoproteins
    • Fatty Acids
    • Synthesis
    • Signal Transduction
    • Metabolism
    • Lipid metabolism
    • Cardiovascular
    • Lipids / Lipoproteins
    • Fatty Acids
    • Metabolism
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Fatty Acid Assays
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Lipid Metabolism Kits
    • Fatty acids
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Lipid Metabolism Kits
    • Obesity
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Fatty acids
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Fatty acid oxidation
  • Relevance

    Fatty acids are carboxylic acids with long hydrocarbon chains. The hydrocarbon chain length may vary from 8-30 carbons. Fatty acids play very important roles in normal metabolism and many disease development. They are precursors to a number of bioactive classes of compounds such as prostaglandins, leucotrienes and others, and have been implicated in diverse functions such as autism, immune system and inflammation response.
  • Alternative names

    • FFA

Associated products

  • Related Products

    • Triglyceride Assay Kit - Quantification (ab65336)
    • Free Glycerol Assay Kit (ab65337)

Images

  • Functional Studies - Free Fatty Acid Assay Kit - Quantification (ab65341)
    Functional Studies - Free Fatty Acid Assay Kit - Quantification (ab65341)Tian, Xiaoxiang et al., PloS one vol. 12,5 e0176873., Fig 6, doi:10.1371/journal.pone.0176873

    Lipolysis were evaluated by measuring free fatty acids released into the culture medium of siCreg1 or si-Scramble 3T3-L1 cells using ab65341, which were treated with BMS-345541 (5 µmol/L), an NF-κB pathway inhibitor, or vehicle (DMSO) for 48 hours prior to lipolysis assessment. ns, no significance; and ***P<0.001.

  • Functional studies - ab65341
    Functional studies - ab65341Samuel P et al., PLoS One 8(1), Fig 7a. DOI: 10.1371/journal.pone.0048425. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

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

  • Func S-1
    Func S-1

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

  • ab65341 Free Fatty Acid Quantification Kit
    ab65341 Free Fatty Acid Quantification Kit

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

  • ab65341 Free Fatty Acid Quantification Kit
    ab65341 Free Fatty Acid Quantification Kit

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

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (242)

Publishing research using ab65341? Please let us know so that we can cite the reference in this datasheet.

ab65341 has been referenced in 242 publications.

  • Maier G  et al. Transcriptomic, proteomic and phosphoproteomic underpinnings of daily exercise performance and zeitgeber activity of training in mouse muscle. J Physiol 600:769-796 (2022). PubMed: 34142717
  • Bhat N  et al. Dyrk1b promotes hepatic lipogenesis by bypassing canonical insulin signaling and directly activating mTORC2 in mice. J Clin Invest 132:N/A (2022). PubMed: 34855620
  • Pham DV & Park PH Adiponectin triggers breast cancer cell death via fatty acid metabolic reprogramming. J Exp Clin Cancer Res 41:9 (2022). PubMed: 34986886
  • Xu Z  et al. Intermittent Fasting Improves High-Fat Diet-Induced Obesity Cardiomyopathy via Alleviating Lipid Deposition and Apoptosis and Decreasing m6A Methylation in the Heart. Nutrients 14:N/A (2022). PubMed: 35057432
  • Dai B  et al. Macrophages in epididymal adipose tissue secrete osteopontin to regulate bone homeostasis. Nat Commun 13:427 (2022). PubMed: 35058428
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-10 of 34 Abreviews or Q&A

Validation of FFA quantification kit for yellow-bellied marmots

Excellent Good 4/5 (Ease of Use)
Abreviews
Abreviews
abreview image
Plasma from yellow-bellied marmots (Marmota flaviventris) was processed according the protocol. A serial dilution of the plasma (shown) was compared to the standard curved (also shown) to determine the optimal dilution factor for subsequent assays.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jul 20 2016

Question

Does tissue need to be fresh or can we snap freeze in liquid nitrogen and grind a small amount prior to homogenising in chloroform/Tx100?


Read More

Abcam community

Verified customer

Asked on Mar 19 2014

Answer


Although we would suggest to use fresh tissue preferably, tissue that is snap-frozen in liquid nitrogen can be used too. The tissue can be ground and then lipid extraction can be done with choloroform-triton.

Read More

Sam Washer

Abcam Scientific Support

Answered on Mar 19 2014

Question

Does the kit detect triglycerides or other molecules that have a fatty acid component?

Read More

Abcam community

Verified customer

Asked on Apr 23 2015

Answer

No, the kit does not detect triglycerides or other molecules that have a fatty acid components. The triglycerides need an enzyme to convert them to free fatty acids and this is not present in any of the kit components.

Read More

Abcam Scientific Support

Answered on Apr 23 2015

Question

Why is the assay not compatible with heparinized samples?

Read More

Abcam community

Verified customer

Asked on Oct 15 2014

Answer

The reason heparinized plasma is not recommended for the free fatty acid assay is because heparin releases lipolytic enzymes that lead to an overestimation of FFA.

Read More

Tom Ruyle

Abcam Scientific Support

Answered on Oct 15 2014

Question

How would I account for endogenous long-chain Acyl-CoAs that are already in my tissue extract samples?

Read More

Abcam community

Verified customer

Asked on Jun 24 2014

Answer

You can test for endogoenous Acyl-CoAs in your samples by including sample wells without added ACS Reagent. In these wells, the long-chain free fatty acids will not be converted to coenzyme A, so if there are any interfering compounds including endogenous Acyl-CoAs, you can subtract this background from the +ACS Reagent wells.

Read More

Caitlin Valued Customer

Abcam Scientific Support

Answered on Jun 24 2014

Question

Is it okay to use this kit with store human serum (-80C)?

Read More

Abcam community

Verified customer

Asked on Nov 26 2013

Answer

We would recommend fresh serum samples for best results. If fresh samples is not an option storage at -80 °C is less likely to degrade the sample quality.

For this product ab65341, blood needs to be collected using an anticoagulant such as EDTA, sodium citrate, sodium fluoride, or ammonium oxalate. Heparinized plasma is not the best choice as heparin could interfere with the assay.

Read More

Tanya Bagrij

Abcam Scientific Support

Answered on Nov 26 2013

Question

I am interested in the Free Fatty Acid Quantification Kit (ab65341). It says to be suitable for the plasma. My question is if this kit suitable for the mouse plasma containing heparin?

Read More

Abcam community

Verified customer

Asked on Oct 31 2013

Answer

Blood needs to be collected using an anticoagulant such as EDTA, sodium citrate, sodium fluoride, or ammonium oxalate. Heparinized plasma is not the best choice as heparin could interfere with the assay.

Read More

Tom Ruyle

Abcam Scientific Support

Answered on Oct 31 2013

Question

What is the pH of the Assay Buffer in the kit?

Read More

Abcam community

Verified customer

Asked on Oct 02 2013

Answer

The details of the assay buffer is considered proprietary information. I can give you a pH range. It is between 7.5-8.2

Read More

Heather Allen

Abcam Scientific Support

Answered on Oct 02 2013

Question






Will it be suitable for lipoxin quantification?

Read More

Abcam community

Verified customer

Asked on May 30 2013

Answer


This kit is good for C-8 or longer fatty acids so it should be suitable for measuring lipoxins.

Read More

Abcam Scientific Support

Answered on May 30 2013

Question

Is C-7 fatty acid suitable for this kit?

Read More

Abcam community

Verified customer

Asked on Apr 26 2013

Answer



Our scientists tried this assay with C7 and C6 fatty acids, but found that the final efficiency of detection decreases by ˜50% in that case.

Therefore I can confirm that the minimal length of suitable fatty acid is C8.

Read More

Abcam Scientific Support

Answered on Apr 26 2013

1-10 of 34 Abreviews or Q&A

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