
New LysoLiveTM Metabolic Lysosomal Enzyme Probes
Marker Gene introduces our new LysoLiveTM Lysosomal Enzyme Activity Kits. Lysosomes are acidic cytoplasmic organelles that are present in all nucleated mammalian cells. Lysosomes have been found to be involved in a variety of cellular processes including repair of the plasma membrane, defense against pathogens, cholesterol homeostasis, bone remodeling, metabolism, apoptosis and cell signaling. Defects in lysosomal enzyme activity have been associated with a variety of diseases including Parkinson’s, Tay-Sachs, Sandhoff, Krabbe and Gaucher’s syndromes. Marker Gene has developed a set of new lysosomal staining compounds that are useful for labeling lysosomes in a live-cell format and are capable of monitoring lysosomal metabolic activity. These new targeted substrates are based upon fluorescent probes that have a low pKa value for optimum fluorescence at the lower physiological pH values found in the lysosomes as well as targeting groups to direct their accumulation to the lysosomes using a live-cell staining format. Two kits are now available: namely the MarkerGeneTM LysoLiveTM Lysosomal Phosphatase Assay Kit (M1376) and the MarkerGeneTM LysoLiveTM Lysosomal Sulfatase Assay Kit (M1377). Several other kits for specific lysosomal enzyme activities will be available soon. For more information about these new assays, please contact our technical assistance department at techservice@markergene.com or see the references below.
- Naleway JJ, Coleman DJ, Guzikowski AP, Williamson SP, " Metabolic Lysosomal Enzyme Probes" FASEB J. 22(1): 1059.1 (2008).
- Carpenter AE,Jones TR, Lamprecht MR, Clarke C, Kang IH, Friman O, Guertin DA, Chang JH, Lindquist RA, Moffat J, Golland P, Sabatini DM (2006) “CellProfiler: image analysis software for identifying and quantifying cell phenotypes.” Genome Biology 7:R100.
- Karageorgos LE, Isaac EL, Brooks DA, Ravenscroft EM, Davey R, Hopwood JJ, Meikle PJ (1997) “Lysosomal Biogenesis in Lysosomal Storage Disorders” Experimental Cell Research 234, 85-97.
- Koenig H (1963) “Vital staining of lysosomes by acridine orange.” J. Cell Biol 19, 87A.
MarkerGeneTM Fluorescent Cellulase Assay Kit

The Fluorescent Cellulase Assay Kit allows fast and easy detection of most cellulases in a microtiter plate based assay format.
Cellulases are a family of enzymes that include ß-Glucosidases, endoglucanases, and exoglucanases. These enzymes cleave the ß-1,4-D-glycosidic bonds that link the glucose units comprising cellulose. In addition to being produced by plants, cellulase activity is found in many fungi and bacteria, including some plant pathogens. Most animal cells are not known to produce cellulase; cellulolytic activity is often carried out in animals by symbionts. However, recent evidence does suggest cellulase production in some animals, such as insects and arthopods. The study of cellulase activity has many applications in plant molecular biology, agriculture, and manufacturing. Cellulase is also becoming important in the development of alternative fuel sources, as glucose obtained from cellulose hydrolysis is easily fermented into ethanol. Activity of most cellulases can be monitored using our long wavelength fluorescent substrate, Resorufin Cellobioside (Product M1238), contained in the kit. Upon cleavage, the fluorescent compound, Resorufin (Product M0202), is released and activity measurements are easily obtained in a microtiter plate based assay format. The kit contains enough substrate for 200 assays and control experiments (100 μL reaction volume) and also contains reference standards and a detailed protocol for use. See the references below for more information and applications.
- Coleman D.J., Studler M.J., Naleway J.J. (2007) "A long-wavelength fluorescent substrate for continuous fluorometric determination of cellulase activity: resorufin-β-D-cellobioside." Anal. Biochem. 371: 146-153
- Shani Z., Dekel M., Roiz L., Horowitz M., Kolosovski N., Lapidot L., Alkan S., Koltai H., Tsabary G., Goren R., Shoseyov O. (2006) "Expression of endo-1,4-β-glucanase (cel1) in Arabidopsis thaliana is associated with plant growth, xylem development and cell wall thickening." Plant Cell Rep, 25: 1067-1074
- Villena G.K., Gutierrez-Correa M. (2006) "Production of cellulase by Aspergillus niger biofilms developed on polyester cloth." Letters in Applied Microbiology. 43: 262-268
- Zhang Q., Bai G., Yang W., Li H., Xiong H. (2006). "Pathogenic cellulase assay of pine wilt disease and immunological localization." Biosci. Biotechnol. Biochem. 70(11): 2727-32
- Nakata T., Miyafuji H., Saka S. (2006) "Bioethanol from cellulose with supercritical water treatment followed by enzymatic hydrolysis." Appl. Biochem. Biotechnol. 129-132: 476-85
- Watanabe H., Tokuda G. (2001) "Animal Cellulases." Cell. Mol. Life Sci. 58: 1167-1178
- "Characterization of a Bifunctional Cellulase and Its Structural Gene." J. Biochem, 270(43): 26012-26019
- Boschker H.T.S., Cappenberg T.E. (1994) "A sensitive method using 4-Methylumbelliferyl-beta-Cellobiose as a Substrate to Measure (1,4)-beta-Glucanase Activity in Sediments." Applied and Environmental Biology, 60(10): 3592-3596
- Chernoglazov V.M., Jafarova A.N., Klyosov A.A. (1989) "Continuous photometric determination of endo-1,4-beta-D-glucanase (cellulase) activity using 4-methylumbelliferyl-beta-D-cellobioside as a substrate." Anal Biochem, 179(1): 186-189
- Thayer D.W. (1978) "Carboxymethylcellulase produced by facultative bacteria from the hind-gut of the termite Reticulitermes hesperus." Journal of General Microbiology, 106(1) 13-8
- Ferrari T., Arnison, P. (1974) "Extraction and Partial Characterization of Cellulases from Expanding Pea Epicotyls." Plant Physiol. 54: 487-493

The Lipase Assay Kit allows fast and easy measurement of lipase activity in vitro, in cell preparations or in vivo using the fluorescent fatty acid substrate 1,2-dioleoyl-3-pyrenyldecanoyl-rac-glycerol (Product M0258).
Lipases are a family of enzymes that release fatty acids from triacylglycerols in a site specific manner. Most lipases have optimum activity for the primary ester groups of triglycerides, while some lipases remove fatty acyl groups from either the C-1 or C-3 acyl positions. The substrate is typically not a single molecule, but a nonaqueous phase of aggregated lipid. Lipase activity, ubiquitous among most cells, can be monitored using the new fluorescent substrate 1,2-dioleoyl-3-pyrenyldecanoyl-rac-glycerol (Product M0258) contained in the kit. Upon cleavage, the fluorescent fatty acid pyrenedecanoic acid (Product M0274) is released and activity measurements are easily obtained either in vitro, in cell preparations, or in vivo. The kit contains enough substrate for numerous assays and control experiments, and also contains reference standards and a detailed protocol for use. See the references below for more information and applications.
- Main, L.A., Okumura-Noji, K., Ohnishi, T., Yokoyama, S. (1998) "Cholesteryl ester transfer protein reaction between plasma lipoproteins." J. Biochem. (Tokyo) 124: 237-243.
- Jaeger, K.E., Ransac, S., Dijkstra, B.W., 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, Q.Q., 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, H.J., 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, A.,D., Shuvaev, V.V., Perova N.V. (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, R.Y., Sawyer, W.H. (1991) "Spectrofluorometric determination of lipase activity." Biochem. Int. 23(5): 825-34.
- Negre, A., Salvayre, R., Dousset, N., Rogalle, P., Dang, Q.Q., Douste-Blazy, L. (1988) "Hydrolysis of fluorescent pyrenetriacylglycerols by lipases from human stomach and gastric juice." Biochim. Biophys. Acta 963: 340-348.
- Hendrickson, H.S. (1994) "Fluorescence-bases assays of lipases, phospholipases, and other lipolytic enzymes." Anal. 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, W.M., Massey, J.B., Gotto, A.M., Pownall, H.J. (1981) "Reassembled Model Lipoproteins." J. Biol. Chem. 256(21): 10815-10819.
- Liodakis, A., Drew, J., Chan, R., Sawyer, W.H. (1991) "Spectrofluorometric determination of lipase activity." Biochem. Intern. 23(5): 825-834.
- Jaeger K.E., Ransac S., Dijkstra B.W., Colson C., van Heuvel M., Misset O., 1994."Bacterial lipases." FEMS Microbiol Rev. 15(1):29-63.
- Arpigny J. L., Jaeger K.-E., "Bacterial lipolytic enzymes: classification and properties. (1999) " Biochem. J. 343: 177-183.
- Kouker G., Jaeger K.E., (1987) "Specific and sensitive plate assay for bacterial lipases." Appl Environ Microbiol) 53(1): 211-3.
- Howard G.T., Vicknair J., MacKie R.I., (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.