Alternative Name: X-Gal Staining Kit
Unit Price (USD):
1 kit $137.50
Bulk Price/unit (when you buy 5 or more): $110.00
Description:
Can be used to monitor and detect enzyme activity in stably or transiently transfected
cells or transgenic tissues using the sensitive chromogenic substrate 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside
(X-Gal).
Note:
The kit contains enough reagents and buffers for up to 100 assays, when run in a microplate assay format, and also includes a detailed protocol for use in detection of β-galactosidase in mammalian cells and tissues.
Application:
The E. coli lacZ gene is
among the most widely used marker genes for mammalian, yeast and bacterial transfection studies due in part to its resistance to intracellular proteolytic degradation. The lacZ gene encodes for the β-Galactosidase (β-Gal) enzyme that catalyzes the hydrolysis of a wide range of β-galactosides. When β-Gal cleaves the glycosidic linkage in X-Gal, a soluble, colorless indoxyl derivative is produced which quickly dimerizes and becomes oxidized to produce a bright blue indigo dye precipitate at the site of β-Gal activity. The dimerization
and oxidation reaction is facilitated, in this kit, by a developer buffer solution containing ferric and ferrous ions. Both transfection efficiency and comparative expression levels can be determined by examining and counting the number of blue cells in the total cell population.
References:
Bout A, Valerio D, Scholte BJ (1993) "In vivo transfer and expression of the lacZ gene in the mouse lung." Exp. Lung Res. 19(2): 193-202
Kobayashi S, Okada M (1993) "A double staining technique using 5-bromo,4-chloro-3-indolyl-beta-D-galactopyranoside (X-gal) and immunoperoxidase in whole Drosophila embryos." Biotech. Histochem. 68(4): 237-9.
Kruger A, Schirrmacher V, von Hoegen P (1994) Scattered micrometastases visualized at the single-cell level: detection and re-isolation of lacZ-labeled metastasized lymphoma cells." Intl. J. Cancer) 58(2): 275-84.
Singh MB, Knox RB (1984) "Quantitative cytochemistry of beta-galactosidase in normal and enzyme deficient (gal) pollen of Brassica campestris: application of the indigogenic method." Histochem 16(12): 1273-96.
Kadokawa Y, Suemori H, Nakatsuji N (1990) "Cell lineage analyses of epithelia and blood vessels in chimeric mouse embryos by use of an embryonic stem cell line expressing the beta-galactosidase gene." Cell Differ. Develop.29(3): 187-94.
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