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Product Details
| Cat # +Size | K721-100 |
|---|---|
| Size | 100 assays |
| Detection Method | Fluorescence (Ex/Em = 482/515 nm) |
| Species Reactivity | Mammalian |
| Applications | Measurement of LPL activity in purified wild type or recombinant protein as well as in plasma, cell and tissue lysates |
| Features & Benefits | • Simple procedure, wide applicability • Fast and convenient • Highly Sensitive Fluorometric assay for measuring Lipoprotein Lipase activity in various biological samples |
| Kit Components | • LPL Assay Buffer• Substrate (in DMSO)• Positive Control • Inhibitor (Orlistat) |
| Storage Conditions | 4°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
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. BioVision’s LPL Activity Assay Kit contains a quenched substrate that fluoresces upon hydrolysis by LPL. The fluorometric intensity is directly proportional to the amount of substrate hydrolyzed. This 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.
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Can you use RIPA buffer to prepare the cell lysates?Do they need to add protease inhibitor to RIPA buffer? May protease inhibitor interfere with this assay?
RIPA buffer is not recommended because the SDS in the buffer tends to inhibit the enzyme that we are trying to assay. They can prepare the lysates in the assay buffer and add protease inhibitor in that. Protease inhibitors will not interfere with the assay.
Is the LPL activity assay specific for LPL or does it also detects HL activity? In our case, a specificity for LPL is very important because we only want to detect LPL activity.
No the LPL activity assay is not specific for LPL. It can also detect Hepatic lipase (HL) activity.
Can we use frozen samples with this assay?
We would recommend using fresh samples. However, frozen samples can be used too provided they have not undergone multiple freeze thaw cycles.
Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
What is the shelf life of this kit?
BioVisions’s assay kits expire 6-12 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet.
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Rodriguez, Susana et al. (2016) Loss of CTRP1 disrupts glucose and lipid homeostasis, Am J Physiol Endocrinol Metab. 2016 Oct 1;311(4):E678-E697.
For more citations of this product click here
