製品の概要

  • 製品名
    Triglyceride Assay Kit - Quantification 
    Triglyceride キット 製品一覧
  • 検出方法
    Colorimetric/Fluorometric
  • サンプルの種類
    Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate
  • アッセイタイプ
    Quantitative
  • 検出感度
    > 2 µM
  • 検出範囲
    2 µM - 10000 µM
  • 全工程の試験時間
    1h 20m
  • 製品の概要

    Triglyceride Assay Kit (ab65336) is a sensitive, easy assay to measure triglyceride concentration in mammalian samples. In the triglyceride assay protocol, triglycerides are converted to free fatty acids and glycerol. Glycerol is then oxidized to generate a product which reacts with a probe to generate color (spectrophotometry at λ= 570 nm) and fluorescence (Ex/Em = 535/587 nm).


    Triglyceride assay protocol summary: 
    - add samples and standards to wells
    - add assay buffer and lipase, and incubate for 20 min
    - add triglyceride reaction mix and incubate for 60 min
    - analyze with microplate reader


    If your sample contains reducing substances such as glucose, fructose or lactose, they are likely to interfere with the assay. In this case, we recommend using Triglyceride Assay Kit (Fluorometric, Reducing samples) ab178780.


    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.

  • 特記事項

    Some components of this kit are available to purchase as standalone reagents if you need more quantities (lipase, enzyme mix, triglyceride standard or triglyceride probe). Please contact our team for more details.

  • 試験プラットフォーム
    Microplate reader

製品の特性

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  • Hepatic triglyceride levels was measured using ab65336 in male and female wild-type (WT) or AT2KO (knockout) mice with either normal diet (ND) or high fat diet (HFD). 

  • Fluorometric triglyceride standard curve: mean of duplicates (+/- SD) with background reads subtracted

  • Triglyceride measured in cell culture lysates showing quantity (nmol) per 1 mln cells.

    Samples with the concentration of 1e7 cells/mL were used. Samples were diluted 40-80 fold and measured fluorometrically.

    HepG2 cells were treated with 25 uM Chloroquine for 72h.

  • Colorimetric triglyceride standard curve using ab65336.

プロトコール

参考文献

This product has been referenced in:
  • García-Ruiz I  et al. Omentectomy Prevents Metabolic Syndrome By Reducing Appetite and Body Weight In A Diet-Induced Obesity Rat Model. Sci Rep 8:1540 (2018). WB . Read more (PubMed: 29367725) »
  • Fernandes GW  et al. The Foxo1-Inducible Transcriptional Repressor Zfp125 Causes Hepatic Steatosis and Hypercholesterolemia. Cell Rep 22:523-534 (2018). Read more (PubMed: 29320745) »
See all 91 Publications for this product

レビューと Q&A

1-10 of 28 Abreviews or Q&A

Question
Answer

We prefer NP40 to Triton or tween in this assay. NP-40 works better to keep lipids in solution and does not create background in the assay.

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Question
Answer

The picoprobe allows ab178780 to be more sensitive than the ab65336. ab178780 detects triglycerides <0.4uM whereas ab65336 can detect 2uM-10mM triglycerides.

I hope this helps!

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Question
Answer

The lipase is from a bacterial source and the standards are from plant oils.

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Question
Answer

Yes, I can confirm that there should be no problem to treat the plasma samples in the same way as the serum, directly applying it.

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Answer

1) The insoluble top layer may contain the triglyceride you wish to measure so please make sure to solubilize as much as possible (without losing any material) and to follow the recommendations (ie. repeated heating steps).
2) SV is indeed the abbreviation for sample volume. For calculating the final results, you need to take into account all the dilution steps you have made during the procedure so dilutions factors (10 and 60 fold need to be included in the calculation).

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Answer

The 96 well plate is not provided with the kit. Please use a black plate (clear bottom) for fluorescence and clear plate for colorimetric analysis.

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Question
Answer

The standard provided with ab65336 has a molecular weight if 885.43 g/mol. 

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Answer

1 mol/L is represented as 1mM, E.g. 50 mM is 50 mmol/L.

Divide by the molecular weight of the substance (average triglyceride molecular weight in your sample) to convert from moles to grams. Then mM/mol weight can be converted to get mg/L.

For this assay, since it is difficult to say the molar mass of triglyceride, unless which specific one is present in the sample is known, the accepted most common way of reporting the data is as mM or µmol/ml.

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Triglycerides from mouse plasma

Good Excellent 5/5 (Ease of Use)
Abreviews
Mouse blood was obtained by cardiac puncture in heparinized tubes and centrifuged at 13000 rpm for 2 min at 4 degrees. Plasma was subsequently diluted 1:20 in PBS for testing. This kit is easy to use and fast, and gives blood triglyceride concentrations in line with what is reported by literature.
Our experience gives reproducible results across the different kit lots.

Abcam user community

Verified customer

投稿 Sep 03 2018

Abreviews
The protocol is easy to understand and perform.
We determined the TG level in liver tissue to use in subsequent assays.

Ms. Ji Hye Han

Verified customer

投稿 Jul 30 2018

1-10 of 28 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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