Unit Price (USD):
1 kit $190.50
Bulk Price/unit (when you buy 5 or more): $152.40
Description:
Allows highly sensitive detection of β-galactosidase activity in mammalian, yeast, bacterial, or plant cells, at a shorter wavelength (blue fluorescence) than other kits. This assay is especially amenable to co-staining with a green (fluorescein, GFP) labeling for dual labeling experiments.
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 beta-galactosidase in vivo. Since this enzyme is generally absent in normal mammalian, yeast, some bacterial and even plant cells, it 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 beta-galactosidase per cell by FACS analysis. Although chromogenic assays of beta-galactosidase activity (i.e. X-Gal) have use, application of the fluorogenic substrate 3-carboxyumbelliferyl β-D-galactopyranoside (CUG) combined with Fluorescence Activated Cell Sorting (FACS) analysis has been shown to be several orders of magnitude more sensitive. In addition, because of its high water solubility and detection limits, the CUG substrate has found extensive use in automated ELISA type assay systems. This assay is especially useful when co-staining with a fluorescein-type cell label (FITC-antibody, fluorescein based substrate, etc.) for dual wavelength detection. Many new instruments have multi-color FACS detection capabilities (see the BD LSRII system for example), that can utilize this new kit. Please consult your instrument manufacturer brochure for more information.
The FACS Blue lacZ β-Galactosidase Detection kit provides all the reagents and a detailed protocol to perform up to 500 automated (10 x 96 well microtiterplate) assays.
References:
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. Guo W, Wu H, (2008) "Detection of LacZ expression by FACS-Gal analysis" Nature Protocols DOI: 10.1038/nprot.2008.163 http://www.natureprotocols.com/2008/08/06/detection_of_lacz_expression_b.php
Keywords:
FACS Fluorescent Blue lacZ β-Galactosidase Detection Kit, FACS, Fluorescent Blue, Fluorescent, lacZ, β-Galactosidase, Beta-Galactosidase, B-Galactosidase, Galactosidase, Detection Kit, Marker Gene
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