Supplied by Roar Biomedical, Inc.
Product No. MAK109
Storage Temperature 2–8 °C
The kit is sufficient for 100 assays in 200 µL total assay volume.
This product is for R&D use only, not for drug, household, or other uses. Please consult the Safety Data Sheet for information regarding hazards and safe handling practices.
Briefly centrifuge vials before opening.
Substrate Emulsion – Vortex before use.
The kit is shipped on wet ice. Storage at 2–8 °C, protected from light, is recommended. DO NOT FREEZE.
Components are stable for 1 year, if stored properly.
All samples and standards should be run in duplicate.
Note: Standards are not to be incubated with samples, use a separate plate for assay of standards.
Note: The substrate is in a liquid crystalline state and difficult to pipette accurately in small volumes. It is suggested to pipette the substrate using a wide barrel pipette tip, or by cutting the end off of a standard pipette tip.
Notes: For unknown samples, it is suggested to test several sample dilutions to ensure the readings are within the linear range of the standard curve.
The amount of LPL to use in the assay and the incubation temperature will depend on the specific activity of the protein. For example, a purified sample of LPL with high specific activity may require dilution with LPL Assay Buffer and/or incubation at 25 °C in order for the assay to provide a linear response. It is recommended first time users start with a lower temperature, ~15 minute incubation time points, and multiple reads to work out a set of conditions.
The steps to validate the assay were adapted from a published procedure,1 using Orlistat (Product No. O4139), also known as tetrahydrolipstatin (THL), to inhibit LPL activity.
Note: LPL as an ammonium sulfate suspension (Product No. L2254) will NOT be inhibited by orlistat. Other sources of LPL without ammonium sulfate, including bacterial LPL (Product No. 62335), work well to validate the assay with inhibitor.
Figure 1.LPL Activity Assay Inhibition with Orlistat
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