Overview

  • Product name

    Lipoprotein Lipase Assay Kit (Fluorometric)
  • Detection method

    Fluorescent
  • Sample type

    Plasma, Cell Lysate, Purified protein, Tissue Lysate
  • Assay type

    Enzyme activity
  • Assay time

    0h 20m
  • Species reactivity

    Reacts with: Other species, Mammals
  • Product overview

    Lipoprotein Lipase Activity Assay Kit (Fluorometric) ab204721 contains a quenched substrate that fluoresces upon hydrolysis by lipoprotein lipase (LPL). The fluorometric intensity is directly proportional to the amount of substrate hydrolyzed.


    This lipase assay detects total lipase activity when no inhibitor is used. Comparing results in the presence or absence of an LPL inhibitor allows for quantification of LPL activity specifically.


    Our results indicate that the majority (~90%) of lipase activity detected by this kit in post-heparin treated mouse plasma is from LPL. To determine the exact LPL specific activity in mouse plasma, measure activity in pre- and post-heparin treated plasma.

  • Notes

    Lipoprotein lipase (LPL) is a member of the lipase family that hydrolyzes triglycerides in chylomicrons and very low-density lipoprotein (VLDL). Digestion of triglycerides in VLDL by LPL leads to their conversion to intermediate-density lipoprotein (IDL) and then low-density lipoprotein (LDL). LPL is found attached to the luminal surface of endothelial cells in the heart, muscle, and adipose tissue. Mutations in lipoprotein lipase can lead to a variety of disorders such as lipoprotein metabolism, hypertriglyceridemia etc. Overexpression of LPL in mice has been shown to promote obesity and insulin resistance.

  • Platform

    Microplate reader

Properties

Images

  • Typical LPL Substrate Standard Curve.

  • Measurement of LPL activity in purified enzyme from Pseudomonas sp. (5 ng), post-heparin treated mouse plasma (2 µl), 7-day post-differentiated 3T3-L1 cell lysate (100 µg), and rat heart lysate (200 µg)

  • Inhibition of LPL activity from post-heparin treated mouse plasma by Angptl 4, an LPL specific inhibitor. The assay was run for 1 hour and the activity was determined by calculating the slope. The IC50 was determined to be 22.6 nM.

  • Inhibition of Positive Control by Orlistat, a generic lipase inhibitor. The assay was run for 1 hour and the IC50 was determined to be 11.4 µM.

Protocols

References

This product has been referenced in:

  • Hiel S  et al. Inulin Improves Postprandial Hypertriglyceridemia by Modulating Gene Expression in the Small Intestine. Nutrients 10:N/A (2018). Read more (PubMed: 29693598) »
See 1 Publication for this product

Customer reviews and Q&As

Answer

The diluted "substrate" is used as the standard in this kit to generate a standard curve of fluorescence intensity based on amount of substrate available for consistent amount of LPL Reaction mix (positive control + ddH2O) to digest. Thus, since active lipoprotein lipase will digest the same substrate compound during the kit reaction, the standard curve can be used to calculate the amount of substrate produced during the reaction time for each sample. As explained in the calculations section, this can then be converted to amound of LPL activity in each sample.

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