MarkerGene™ Long Wavelength Fluorescent Lipase Assay Kit
Product ID: M1214
This kit allows fast and easy measurement of lipase activity in vitro, in cell preparations or in vivo using the long wavelength substrate resorufin oleate.
- Henkel M, Schmidberger A, Vogelbacher M, Kühnert C, Beuker J, Bernard T, Schwartz T, Syldatk C, Hausmann R. (2014) "Kinetic modeling of rhamnolipid production by Pseudomonas aeruginosa PAO1 including cell density-dependent regulation." Appl Microbiol Biotechnol. 98(16):7013-25.
- Main LA, Okumura-Noji K, Ohnish, T, Yokoyama S. (1998) "Cholesteryl ester transfer protein reaction between plasma lipoproteins." J. Biochem. (Tokyo) 124: 237-243.
- Jaeger KE, Ransac S, Dijkstra BW, Colson C, van Heuvel M, Misset O. (1994) "Bacterial Lipases" FEMS Microbiol. Rev. 15(1): 29-63.
- Dousset N, Negre A, Salvayre R, Rogalle P, Dang QQ, Douste-Blazy L. (1988) "Use of a fluorescent radiolabeled triacylglycerol as a substrate for lipoprotein lipase and hepatic triglyceride lipase." Lipids 23: 605-608.
- Galla HJ, Theilen U, Hartmann W. (1979) "Transversal Mobility in Bilayer Membrane Vesicles: Use of Pyrene Lecithin as Optical Probe" Chem Phys Lipids 23: 239-251.
- Dergunov AD, Shuvaev VV, Perova NV. (1989) "Topo-dynamic characteristics of human plasma VLDL apolipoproteins and efficiency of triacylglycerol hydrolysis by lipoprotein lipase." Biochim Biophys Acta 1005(1): 79-86.
- Liodakis A, Drew J, Chan RY, Sawyer WH. (1991) "Spectrofluorometric determination of lipase activity." Biochem Int 23(5): 825-34.
- Negre A, Salvayre R, Dousset N, Rogalle P, Dang QQ, Douste-Blazy L. (1988) "Hydrolysis of fluorescent pyrenetriacylglycerols by lipases from human stomach and gastric juice." Biochim Biophys Acta 963: 340-348.
- Hendrickson HS. (1994) "Fluorescence-bases assays of lipases, phospholipases, and other lipolytic enzymes." Analyt Biochem 219: 1-8.
- Negre-Salvayre A, Abouakil N, Lombardo D, Salvayre R. (1990) "Hydrolysis of fluorescent pyrene-acyl esters by human pancreatic carboxylic ester hydrolase and bile salt-stimulated lipase." Lipids 25(8): 428-434.
- Negre A, Maret A, Douste-Blazy L, Gatt S, Salvayre R. (1988) "Relative fluorescence of normal and acid lipase-deficient cultured fibroblasts following administration of pyrene decanoic acid." Biochim Biophys Acta 960: 401-409.
- Negre A, Salvayre R, Dagan A, Gatt S. (1985) "New fluorometric assay of lysosomal acid lipase and its application to the diagnosis of Wolman and cholesteryl ester storage diseases." Clin Chim Acta 149: 81-88.
- Rosseneu M, Taveirne M, Caster H, Van Biervliet J. (1985) "Hydrolysis of very-low-density lipoproteins labeled with a fluorescent triacylglycerol: 1,3-dioleoyl-2-(4-pyrenylbutanoyl)glycerol." Eur J Biochem 152: 195-198.
- Mantulin WM, Massey JB, Gotto AM, Pownall HJ. (1981) "Reassembled Model Lipoproteins." J Biol Chem 256(21): 10815-10819.
- Liodakis A, Drew J, Chan R, Sawyer WH. (1991) "Spectrofluorometric determination of lipase activity." Biochem Intern 23(5): 825-834.
- Arpigny J L, Jaeger KE.(1999) "Bacterial lipolytic enzymes: classification and properties." Biochem J 343: 177-183.
- Kouker G, Jaeger KE. (1987) "Specific and sensitive plate assay for bacterial lipases." Appl Environ Microbiol 53(1): 211-3.
- Howard GT, Vicknair J, MacKie RI,. (2001) "Sensitive plate assay for screening and detection of bacterial polyurethanase activity." Lett Appl Microbiol 32(3): 211-4.
- Salvayre R, Negre A, Radom J, Douste-Blazy L. (1986) "Fluorometric Assay for Pancreatic Lipase" Clin Chem 32(8): 1532-1536.
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But be aware that the tissues may have multiple lipase enzymes present. The different isoforms of the lipases could have different Vmax values. And the crude cell lysate may also mask the lipase activity as well. Using the kit with cell lysates will therefore only tell you that the lipase activity in cell line 1 is say 50% of that in cell line 2, without doing some other purification. You can do this by either plotting the kinetics, as you mention, or measuring at a single time point for all the samples, i.e. 20 min. or 60 min.
The resorufin standard is used to convert the RFU values from the plate reader or fluorometer into actual concentrations of substrate turnover. You don't really need a standard curve for this, you can just use one point, but it is good to see what the linear range is for your instrument. If your enzyme assay at a certain time period gives a RFU reading of, say 5000, then you can use the curve to calculate the concentration of resorufin that gives the same RFU signal. This will then correlate to the concentration of resorufin generated in the assay and (by definition) the concentration of substrate that has been used by the enzyme at that point. This allows you to calculate velocities (umole/min/mL) from the RFU data.
- Arpigny JL, Jaeger KE (1999)"Bacterial lipolytic enzymes: classification and properties." Biochem. J. 343: 177-183.
- Kouker G, Jaeger KE (1987) "Specific and sensitive plate assay for bacterial lipases." Appl Environ Microbiol 53(1): 211-3.
- Howard GT, Vicknair J, MacKie RI (2001) "Sensitive plate assay for screening and detection of bacterial polyurethanase activity." Lett. Appl. Microbiol. 32(3): 211-4.
- Jaeger KE, Ransac S, Dijkstra BW, Colson C, van Heuvel M, Misset O (1994) "Bacterial lipases." FEMS Microbiol Rev. 15(1):29-63.