QuantiChrom™ Lipase Assay Kit

Protocol SDS

Application

• For quantitative determination of lipase enzyme activity.

Key Features

• Sensitive and accurate. Linear detection ranges from 40 to 1600 U/L lipase activity in a 96-well plate assay.
• Convenient and high throughput. The procedure involves adding a single working reagent and reading the optical density at 10 min and 20 min at room temperature or 37°C. Can be automated to process thousands of samples per day.

Method

• OD412nm

Samples

• Serum, plasma, saliva, urine, etc

Species

• All

Procedure

• 20 min

Size

• 100 tests

Detection Limit

• 40 U/L

Shelf Life

• 12 months

More Details

LIPASE catalyzes the hydrolysis of ester bonds on the glycerol backbone of a lipid substrate. In humans, pancreatic lipase is the key enzyme responsible for breaking down fats in the digestive system by converting triglycerides to monoglycerides and free fatty acids. Human pancreatic lipase and its related protein 2 are the main lipases secreted by the pancreas. In acute pancreatitis, lipase levels can rise 5 to 10-fold within 24 to 48 hours. Increased activities have also been associated with pancreatic duct obstruction, pancreatic cancer, kidney disease, salivary gland inflammation, bowel obstruction, and other pancreatic diseases. Decreased levels may indicate permanent damage to lipase-producing cells in the pancreas. Simple, direct, and automation-ready procedures for measuring lipase activity are very desirable. BioAssay Systems QuantiChrom™ Lipase Assay is based on an improved dimercaptopropanol tributyrate (BALB) method, in which SH groups formed from lipase cleavage of BALB react with 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB) to form a yellow colored product. The color intensity, measured at 412 nm, is proportionate to the enzyme activity in the sample.

Can this kit be used to measure lipase activity in cooking oil?

No, we tested it with one oil sample (Sicilian Selezione Extra Virgin Olive Oil). We mixed 600 µL WR with 400 µL oil (control 600 µL WR alone) and shaked for about 4 hrs. We did not see difference in color between control and sample.

Can this kit be used to determine phospholipase activity (phospholipase C)?

Yes, if you use purified phospholipase (e.g.by immunoprecipitation)

Pitino, MA et al (2019). Is frozen human milk that is refused by mother?s own infant suitable for human milk bank donation? Breastfeeding Medicine: The Official Journal of the Academy of Breastfeeding Medicine, 14(4), 271-275. Assay: Lipase in human milk.

Imanikia, S et al (2019). Xbp-1 remodels lipid metabolism to extend longevity. Cell Reports, 28(3), 581-589.e4. Assay: Lipase in nematode lysates.

Panee, J et al (2020). Chronic marijuana use moderates the correlations of serum cholesterol with systemic mitochondrial function and fluid cognition. Mitochondrion, 52, 135-143. Assay: Lipase in human serum.

Li, L et al (2020). Effects of protocatechuic acid on ameliorating lipid profiles and cardio-protection against coronary artery disease in high fat and fructose diet fed in rats. Journal of Veterinary Medical Science, 82(9), 1387-1394. Assay: Lipase in rat serum.

Fassbinder-Orth, C. A., Killpack, T. L., Goto, D. S., Rainwater, E. L., & Shearn-Bochsler, V. I. (2018). High costs of infection: Alphavirus infection reduces digestive function and bone and feather growth in nestling house sparrows (Passer domesticus). PloS one, 13(4), e0195467. Assay: Lipase in sparrow liver.

Ranawade, A., Mallick, A., & Gupta, B. P. (2018). PRY-1/Axin signaling regulates lipid metabolism in Caenorhabditis elegans. PloS one, 13(11), e0206540. Assay: Lipase in worm tissue.

Srihardyastutie, A., & Soeatmadji, D. W. (2018). The Proteomic Analysis of Pancreatic Exocrine Insufficiency Protein Marker in Type 2 Diabetes Mellitus Patients. In IOP Conference Series: Materials Science and Engineering 299(1) p 012021. Assay: Lipase in human blood.

Li, Y., Nguyen, D. N., de Waard, M., Christensen, L., Zhou, P., Jiang, P. & Dalsgaard, T. K. (2017). Pasteurization procedures for donor human milk affect body growth, intestinal structure, and resistance against bacterial infections in preterm pigs. The Journal of nutrition, 147(6), 1121-1130. Assay: Lipase in human milk.

Shen, A., Kim, H. J., Oh, G. S., Lee, S. B., Lee, S. H., Pandit, A. & Cho, E. Y. (2017). NAD+ augmentation ameliorates acute pancreatitis through regulation of inflammasome signalling. Scientific reports, 7(1), 3006. Assay: Lipase in mice serum.

Flanagan, J. L., Khandekar, N., Zhu, H., Watanabe, K., Markoulli, M., Flanagan, J. T., & Papas, E. (2016). Glycerol monolaurate inhibits lipase production by clinical ocular isolates without affecting bacterial cell viability. Investigative ophthalmology & visual science, 57(2), 544-550. Assay: Lipase in bacteria cells.

Molinari, C.E., et al. (2011). The effect of storage at 25 deg C on proteins in human milk. International Dairy Journal 21(4):286-293. Assay: Lipase in human milk.

Tonsberg, H., et al. (2011). Effect of bile on the oral absorption of halofantrine in polyethylene glycol 400 and polysorbate 80 formulations dosed to bile duct cannulated rats. J Pharm Pharmacol 63(6):817-24. Assay: Lipase in rat intestine.

Freitag, T.L., et al. (2010). human risk allele HLA-DRB1*0405 predisposes class II transgenic Ab0 NOD mice to autoimmune pancreatitis. Gastroenterology 139(1):281-91. Assay: Lipase in mouse serum.

Hu, M., et al. (2010). Effect of prolonged starvation on body weight and blood-chemistry in two horseshoe crab species: Tachypleus tridentatus and Carcinoscorpius rotundicauda (Chelicerata: Xiphosura). Journal of Experimental Marine Biology and Ecology 395(1-2):112-9. Assay: Lipase in horseshoe crab blood.

Nachnani, J.S., et al. (2010). Biochemical and histological effects of exendin-4 (exenatide) on the rat pancreas. Diabetologia 53(1):153-9. Assay: Lipase in rat serum.

Hasan, F., et al. (2009). Methods for detection and characterization of lipases: A comprehensive review. Biotechnol Adv 27(6):782-98. Assay: Lipase in microbes.

Mudgil, Y., et al. (2009). Arabidopsis N-MYC DOWNREGULATED-LIKE1, a positive regulator of auxin transport in a G protein-mediated pathway. Plant Cell 21(11):3591-609. Assay: Lipase in Arabidopsis thaliana recombinant proteins.

Narbonne, P., Roy, R. (2009). Caenorhabditis elegans dauers need LKB1/AMPK to ration lipid reserves and ensure long-term survival. Nature 457(7226):210-4. Assay: Lipase in C.elegans larvae.

Qatanani, M., et al. (2009). Macrophage-derived human resistin exacerbates adipose tissue inflammation and insulin resistance in mice. J Clin Invest 119(3):531-9. Assay: Lipase in mouse muscle tissue.

Gabbi, C., et al. (2008). Pancreatic exocrine insufficiency in LXRbeta-/- mice is associated with a reduction in aquaporin-1 expression. Proc Natl Acad Sci U S A 105(39):15052-7. Assay: Lipase in mouse serum.

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If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

– Fast turnaround
– Quality data
– Low cost

Please email or call 1-510-782-9988 x 2 to discuss your projects.