Molecular Analysis of Transcriptional Control: The FACS-Galactosidase Assay
The FACS-galactosidase (FACS-Gal) assay measures Escherichia coli lacZ-encoded ′-galactosidase activity in eukaryotic cells. Based on analysis by flow cytometry, the enzyme assay allows the simultaneous use of even a single, stably integrated lacZ gene as a reportergene for functional analysis of transcriptional control elements. Apart from providing good statistics due to the high number of cells analyzed, lacZ expression can be measured quantitatively on a per cell basis. Also, lacZ expression can be correlated with other cellular parameters such as immunophenotype. Cells can be sorted alive according to lacZ expression, from less than 5 to more than 50 000 enzyme molecules per cell, for further cellular and molecular studies.
KeywordsGalactosidase Activity lacZ Expression Cellular Parameter Staining Medium Hydroxyethylpiperazine Ethanesulfonic Acid
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