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MarkerGeneTM Cellular Senescence Microtiterplate Assay Kit
 Studies have shown that ß-galactosidase histochemical staining at pH 6 is a useful enzymatic marker of cellular senescence (the so-called senescence associated beta-galactosidase; SA-ß-Gal). While cells entering senescence show this enzymatic activity, immortalized cell types, including tumor or other quiescent cells, are not stained under these conditions. The implications of these methods in such diverse areas as age-related pathology research, tumor analysis and tissue culture maintenance is significant. The SA-ß-Gal assay is one of the most widely accepted markers of cell aging. Often analysis is performed using the chromogenic substrate X-Gal. However, the substrate X-Gal has limited sensitivity and is difficult to quantify. The degree of blue staining in positive cells varies both with cell type and staining conditions, and this introduces subjectivity to results. The MarkerGeneTM Cellular Senescence Microtiterplate Assay Kit (M1405) uses the high-sensitivity substrate fluorescein di-ß-D-galactopyranoside (FDG) to quantify senescent cells. FDG turnover produces the highly fluorescent product fluorescein, which is easily detected using an appropriate filter set for fluorescein fluorescence (i.e. EX 485nm and EM 535 nm) in a microtiterplate assay format. This assay ... more
New Red Bioluminescent Luciferase Vectors
Researchers at Marker Gene have developed a new luciferase gene using point mutation and codon optimization of the luciferase sequence from the natural luciferase gene isolated from Luciola cruciata (the Japanese firefly). This novel recombinant DNA has been incorporated into vectors (M1394 and M1395) containing the highly expressing CMV and SV40 promoters for high expression activities in mammalian cells. The gene codes for an improved amino acid sequence which exhibits long-wavelength light emission (red color, EM 619 nm), as well as improved thermostability and higher expression levels in mammalian cells compared with other luciferases including the native luciferase derived from the American firefly Photinus pyralis. The expressed enzyme uses the same substrate, D-Luciferin (M0237), that is used with other luciferases, and activity can easily be detected using our convenient MarkerGeneTM Live Cell Luciferase Assay Kit (M0626). Additional substrates for use in detecting cloned activity in living tissues and in vivo include the membrane permeant D-Luciferin, ethyl ester (M0906). This makes these new vectors ideal candidates for use in vivo detection methods ... more
MarkerGene™ Fluorescent Lipase Assay Kit
 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 (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 (M0258) contained in the kit. Upon cleavage, the fluorescent fatty acid pyrenedecanoic acid (M0274) is released...more
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