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    products/assay-kits/lipoprotein-lipase-assay-kit-fluorometric-ab204721.pdf

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Lipoprotein Lipase Assay Kit (Fluorometric) (ab204721)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (1)References (9)

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Lipoprotein Lipase Activity Assay Kit (Fluorometric) (ab204721)
  • Lipoprotein Lipase Activity Assay Kit (Fluorometric) (ab204721)
  • Lipoprotein Lipase Activity Assay Kit (Fluorometric) (ab204721)
  • Lipoprotein Lipase Activity Assay Kit (Fluorometric) (ab204721)

Key features and details

  • Assay type: Enzyme activity
  • Detection method: Fluorescent
  • Platform: Microplate reader
  • Assay time: 20 min
  • Sample type: Cell Lysate, Plasma, Purified protein, Tissue Lysate

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Overview

  • Product name

    Lipoprotein Lipase Assay Kit (Fluorometric)
    See all Lipoprotein lipase kits
  • Detection method

    Fluorescent
  • Sample type

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

    Enzyme activity
  • Assay time

    0h 20m
  • Species reactivity

    Reacts with: Mammals, Other species
  • 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

    This product is manufactured by BioVision, an Abcam company and was previously called K721 Lipoprotein Lipase Activity Fluorometric Assay Kit. K721-100 is the same size as the 100 test size of ab204721.

    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

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 100 tests
    Inhibitor (Orlistat) 1 x 20µl
    LPL Assay Buffer 1 x 5ml
    Positive Control 1 vial
    Substrate (in DMSO) 1 x 10µl
  • Research areas

    • Cardiovascular
    • Lipids / Lipoproteins
    • Lipid Metabolism
    • Lipases
    • Signal Transduction
    • Metabolism
    • Lipid metabolism
    • Stem Cells
    • Mesenchymal Stem Cells
    • Adipogenesis
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of lipids and lipoproteins
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Lipid metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Lipases
    • Metabolism
    • Types of disease
    • Obesity
  • Function

    The primary function of this lipase is the hydrolysis of triglycerides of circulating chylomicrons and very low density lipoproteins (VLDL). Binding to heparin sulfate proteogylcans at the cell surface is vital to the function. The apolipoprotein, APOC2, acts as a coactivator of LPL activity in the presence of lipids on the luminal surface of vascular endothelium.
  • Involvement in disease

    Defects in LPL are the cause of lipoprotein lipase deficiency (LPL deficiency) [MIM:238600]; also known as familial chylomicronemia or hyperlipoproteinemia type I. LPL deficiency chylomicronemia is a recessive disorder usually manifesting in childhood. On a normal diet, patients often present with abdominal pain, hepatosplenomegaly, lipemia retinalis, eruptive xanthomata, and massive hypertriglyceridemia, sometimes complicated with acute pancreatitis.
  • Sequence similarities

    Belongs to the AB hydrolase superfamily. Lipase family.
    Contains 1 PLAT domain.
  • Post-translational
    modifications

    Tyrosine nitration after lipopolysaccharide (LPS) challenge down-regulates the lipase activity.
  • Cellular localization

    Cell membrane. Secreted. Locates to the plasma membrane of microvilli of hepatocytes with triacyl-glycerol-rich lipoproteins (TRL). Some of the bound LPL is then internalized and located inside non-coated endocytic vesicles.
  • Target information above from: UniProt accession P06858 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • EC 3.1.1
    • EC 3.1.1.34
    • HDLCQ11
    • LIPD
    • LIPL_HUMAN
    • Lipoprotein lipase
    • LPL
    • LPL protein
    • MGC137861
    see all

Images

  • Lipoprotein Lipase Activity Assay Kit (Fluorometric) (ab204721)
    Lipoprotein Lipase Activity Assay Kit (Fluorometric) (ab204721)

    Typical LPL Substrate Standard Curve.

  • Lipoprotein Lipase Activity Assay Kit (Fluorometric) (ab204721)
    Lipoprotein Lipase Activity Assay Kit (Fluorometric) (ab204721)

    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)

  • Lipoprotein Lipase Activity Assay Kit (Fluorometric) (ab204721)
    Lipoprotein Lipase Activity Assay Kit (Fluorometric) (ab204721)

    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.

  • Lipoprotein Lipase Activity Assay Kit (Fluorometric) (ab204721)
    Lipoprotein Lipase Activity Assay Kit (Fluorometric) (ab204721)

    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

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (9)

Publishing research using ab204721? Please let us know so that we can cite the reference in this datasheet.

ab204721 has been referenced in 9 publications.

  • Shi H  et al. CRISPR/Cas9 based blockade of IL-10 signaling impairs lipid and tissue homeostasis to accelerate atherosclerosis. Front Immunol 13:999470 (2022). PubMed: 36110841
  • Gencer S  et al. Adipocyte-Specific ACKR3 Regulates Lipid Levels in Adipose Tissue. Biomedicines 9:N/A (2021). PubMed: 33917642
  • Bugler-Lamb AR  et al. Adipocyte integrin-linked kinase plays a key role in the development of diet-induced adipose insulin resistance in male mice. Mol Metab 49:101197 (2021). PubMed: 33647469
  • Ringel AE  et al. Obesity Shapes Metabolism in the Tumor Microenvironment to Suppress Anti-Tumor Immunity. Cell 183:1848-1866.e26 (2020). PubMed: 33301708
  • Kluge S  et al. Simple and rapid real-time monitoring of LPL activity in vitro. MethodsX 7:100865 (2020). PubMed: 32274337
  • Gao M  et al. ApoC2 deficiency elicits severe hypertriglyceridemia and spontaneous atherosclerosis: A rodent model rescued from neonatal death. Metabolism 109:154296 (2020). PubMed: 32562799
  • Brocker CN  et al. Long non-coding RNA Gm15441 attenuates hepatic inflammasome activation in response to PPARA agonism and fasting. Nat Commun 11:5847 (2020). PubMed: 33203882
  • Kamermans A  et al. Reduced Angiopoietin-Like 4 Expression in Multiple Sclerosis Lesions Facilitates Lipid Uptake by Phagocytes via Modulation of Lipoprotein-Lipase Activity. Front Immunol 10:950 (2019). PubMed: 31130950
  • Hiel S  et al. Inulin Improves Postprandial Hypertriglyceridemia by Modulating Gene Expression in the Small Intestine. Nutrients 10:N/A (2018). PubMed: 29693598

Customer reviews and Q&As

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

Question

I don't see a standard included in the kit content, and in the protocol the standard preparation section is confusing. Can you tell me what I suppose to use as standard?

Read More

Abcam community

Verified customer

Asked on Jan 20 2016

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.

Read More

Kevin Hanson

Abcam Scientific Support

Answered on Jan 20 2016

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