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Extract protein from yeast two-hybrid reporter systems and measure beta-galactosidase activity using compatible Y-PER Reagent and b-Gal Assay Reagent.
| 產品號碼 | Quantity |
|---|---|
| 75768 | 75 mL |
Thermo Scientific Yeast β-Galactosidase Assay Kit is specially formulated for yeast cells and ideal for identifying protein interactions in-vivo using two-hybrid systems.
Features of Yeast beta-Galactosidase Assay Kit:
Yeast β-Galactosidase Assay Kit allows qualitative or quantitative determination of β-galactosidase activity in solution directly from colonies growing on solid medium. A portion of the colony is suspended in a mixture of Thermo Scientific Y-PER Yeast Protein Extraction Reagent and β-Galactosidase Assay Buffer. After a brief incubation period, the solution turns yellow from the hydrolysis of o-nitrophenyl-β-D-galactopyranoside (ONPG) to o-nitrophenol (ONP) and galactose in a mildly alkaline solution. The assay becomes quantitative if calibrated to cell number by measuring the optical density at 660nm.
The gene encoding β-galactosidase (lacZ) of E. coli has been widely used as a reporter gene in many different prokaryotic and eukaryotic organisms. β-Galactosidase activity is also commonly used as an indicator of protein-protein interactions in-vivo using two-hybrid systems. The interaction strength is verified or quantitated using a β-galactosidase activity assay.
The gene encoding β-galactosidase (lacZ) of E. coli has been widely used as a reporter gene in many different prokaryotic and eukaryotic organisms. β-Galactosidase activity is also commonly used as an indicator of protein:protein interactions in vivo using two-hybrid systems. The interaction strength is verified or quantitated using a β-galactosidase activity assay.
Common protocols for determining β-galactosidase activity from liquid cultures involve harvesting cells by centrifugation, washing cells several times, and lysing cells by either subjecting them to freeze/thaw cycles or by treating them with detergents or organic solvents. These traditional protocols are not amenable to projects that involve screening cells growing in 96-well plates. The Yeast β-Galactosidase Assay Kit allows cells obtained from solid media or liquid culture to be assayed directly with no harvesting or washing steps.