Product Details
| abID | ab102524 |
|---|---|
| Cat # +Size | K722-100 |
| Size | 100 assays |
| Detection Method | Absorbance (570 nm) |
| Species Reactivity | Mammalian |
| Applications | This kit detects lipase activity as low as 0.02 mU per well. |
| Features & Benefits | • Simple procedure; takes ~ 60-90 minutes • Fast and convenient • Sensitive assays for measuring Lipase activity in various biological samples. |
| Kit Components | • Lipase Assay Buffer • OxiRed Probe • Dimethylsulfoxide (DMSO) • Enzyme Mix (lyophilized) • Lipase Substrate • Glycerol Standard (100mM) • Lipase Positive Control |
| Storage Conditions | -20°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
Lipases perform essential roles in the digestion, transport and processing of dietary lipids (e.g. fats and oils) in living organisms. In humans, pancreatic lipase is the key enzyme responsible for breaking down fats in the digestive system by converting triglyceride to monoglyceride and free fatty acid. Pancreatic lipase monitoring is also used to help diagnose Crohn's disease, cystic fibrosis and celiac disease. Damage to the pancreas can exhibit a 5-10 fold increase of serum lipase levels within 24 to 48 hours. In BioVision’s Lipase Assay Kit, lipase hydrolyzes a triglyceride substrate to form glycerol which is quantified enzymatically by via monitoring a linked change in the OxiRed probe absorbance (λ=570nm). This assay is rapid, simple, sensitive, and reliable, as well as, suitable for high throughput activity screening of lipase. This kit detects lipase activity as low as 0.02 mU per well.
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How do we collect blood for this assay?
Blood should be collected in heparin tubes and EDTA/metal chelators should be avoided.
My controls without the substrate is showing a high reading. Why?
When you omit the substrate and still get high values from the controls, that means there is comparable amounts of glycerol in the samples. One way to get rid of glycerol is to filter the samples through a molecular cut off filter. The Lipase enzyme mol. wt will allow it to remain in the retentate and glycerol will be in the filtrate for a 10KDa spin column. Use the retentate to do the assay. You can try this for a couple of samples to see if it makes a difference in the control measurements.
There is high values in the blank. What could be the reason?
There could be multiple reasons contributing to a high blank OD. 1. The probe is extremely sensitive to handling. Exposure to air or moisture can degrade the probe. 2. There could be contamination in the assay buffer or other reagetns.
Will this kit detect human hepatic lipase in post-heparin plasma? Or lipoprotein lipase?
K722 will detect generic lipase activity that is the ability of the lipase to convert triglyceride to glycerol. Heparin-treated plasma should work. The following publication addresses variability issues using heparin plasma for lipase assays in general which you might find useful for your experiments: http://www.ncbi.nlm.nih.gov/pubmed/11350025
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.
What is the difference between these two kits?
In K722-100, lipase hydrolyzes a triglyceride substrate to form glycerol, which is quantified enzymatically via monitoring a linked change in the OxiRed probe absorbance (λ = 570nm). In K723-100, lipases hydrolyze a specific substrate to generate a product, which then reacts with the DTNB probe to generate color (λ = 412 nm). Basically, principle for both the kits is the same. However, lipase substrate and detection probe used are different for each of kits.
Tilokwattanothai, A. et al., (2017) Hypoglycemic and hypolipidemic properties of herbal tea on Wistar rat, Chiang Mai Veterinary Journal, 2017, 15(1): 25-35
Anuwat T et al., (2017) Hypoglycemic and hypolipidemic properties of herbal tea on Wistar rat, Chiang Mai Veterinary Journal, 2017, 15(1): 25-35
Cui K et al., (2017) Ontogenetic development of digestive enzymes in larval and juvenile crimson snapper Lutjanus erythopterus (Bloch 1790), Aquaculture Res, 2017, 48(8):4533-4544
Kumar M., Naresh et al. (2016) Molecular characterization of human ABHD2 as TAG lipase and ester hydrolase Biosci Rep. 2016 Jul 15;36(4).
Pruszynska-Oszmalek et al., In ovo injection of prebiotics and synbiotics affects the digestive potency of the pancreas in growing chickens. Poultry Science, Aug 2015; 94: 1909 - 1916.
For more citations of this product click here
