Although chromogenic assays of β-galactosidase activity (i.e. X-Gal) have use, application of the fluorogenic substrate 3-carboxyumbelliferyI β-D-galactopyranoside (CUG) (blue fluorescence) 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. Abcam’s FACS Blue LacZ beta Galactosidase detection kit (ab189815) is especially useful for dual-labeling experiments where the cells to be analyzed are also labeled with a fluorescein based probe (FITC-labeled antibody, fluorescein-based substrate, etc.).
Emission: 460 nm.
Excitation: 390 nm.
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 β-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 β-galactosidase per cell by FACS analysis.
Abcam’s FACS Blue LacZ beta Galactosidase detection kit (ab189815) provides reagents and protocol to perform up to 500 tests (10 x 96 well microtitre plates).
|Inhibitor||1 x 1ml|
|Reference Standard||1 x 0.5ml|
|Substrate Reagent||1 x 0.5ml|
Our Abpromise guarantee covers the use of ab189815 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Other||Use at an assay dependent concentration.
Measures the expression of reporter gene, beta-galactosidase.