Product M0259
MarkerGeneTM in vivo lacZ ß-Galactosidase Intracellular Detection Kit


MarkerGeneTM in vivo lacZ ß-Galactosidase Intracellular Detection Kit

CAS Number: [N/A]

Unit Price (USD): 1 kit $190.50

Bulk Price/unit (when you buy 5 or more): $152.40

Description:
The MarkerGeneTM in vivo lacZ ß-Galactosidase Intracellular Detection Kit allows ultra-sensitive detection of ß-galactosidase activity in live mammalian, yeast, bacterial, or plant cells. Especially useful for FACS analysis.

Application:
One of the most common reporter genes used in molecular biology applications is the E.Coli lacZ gene that codes for an active subunit of ß-galactosidase in vivo. Since this enzyme is generally absent in normal mammalian cells, can be detected at very low levels, and since the enzyme has a wide substrate specificity, monitoring lacZ expression (and therefore co-expressed genes or promoter efficiency) has become routine to the point of detection of as few as 5 copies of ß-galactosidase per cell.

Although chromogenic assays of ß-galactosidase activity (i.e. X-Gal) have use, the recent application of the fluorogenic substrate fluorescein di-ß-D-galactopyranoside (FDG) combined with Fluorescence microscopic analysis (confocal microscopy) analysis has been shown to be several orders of magnitude more sensitive. In addition, because of its improved detection limits, the FDG substrate has found extensive use in automated ELISA type assay systems .

This kit uses the ß-galactoside analog fluorescein di-ß-D-Galactopyranoside (FDG) in a protocol that sensitively distinguishes lacZ+ vs. lacZ- cells. See also Product M0250 for more information and references.

References:

  • "Endothelial-to-mesenchymal transition contributes to cardiac fibrosis." Zeisberg EM, Tarnavski O, Zeisberg M, Dorfman AL, McMullen JR, Gustafsson E, Chandraker A, Yuan X, Pu WT, Roberts AB, Neilson EG, Sayegh MH, Izumo S, Kalluri R. Nature Medicine 13: 952-961 (01 Aug 2007).
  • Roederer M., Fiering S., Herzenberg L.A. (1991) "FACS-Gal: Flow Cytometric Analysis and Sorting of Cells Expressing Reporter Gene Constructs." Methods: Companion to Meth. Enzymol. 2: 248.
  • Nolan G.P., Fiering S., Nicolas J.F., Herzenberg L.A., (1988) "Fluorescence-activated cell analysis and sorting of viable mammalian cells based on beta-D-galactosidase activity after transduction of Escherichia coli lacZ." Proc. Natl. Acad. Sci. U S A 85: 2603-2607.
  • Krasnow M.A., Cumberledge S., Manning G., Herzenberg L.A., Nolan G.P. (1991) "Whole animal cell sorting of Drosophila embryos." Science 251: 81-85.
  • Fiering S., Nolan G., Herzenberg L.A. (1988) "Inhibitors of Endogenous Mammalian B-Galactosidase (B-Gal) or Transduced E. coli B-Gal Improve the Sensitivity and Versatility of the FACS-FDG Fluorogenic Assay for E. coli LacZ Expression in Viable Mammalian Cells." Cytometry (Suppl 2) 1.



    • Keywords:

    β-Galactosidase, b-Galactosidase, b Galactosidase, beta Galactosidase, Galactosidase, B-Galactosidase, Intracellular Detection Kit, Intracellular, Detection Kit, M0259, in vivo, lacZ, ß-Galactosidase, Marker Gene