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Yeast beta-Galactosidase Assay Kit


Efficient cell lysis and sensitive colorimetric detection of beta-galactosidase in yeast cells.

Yeast beta-Galactosidase Assay Kit

The 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.

The Yeast β-Galactosidase Assay Kit allows for 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 the 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.

Highlights:

  • Efficient lysis of yeast cells and a colorimetric detection system
  • Quantitative or qualitative assay for yeast colonies grown on solid media or in suspension
  • Colorimetric assay easily measured in a plate reader or spectrophotometer (420nm)
  • Adaptable to 96-well microplates or tube assay formats
  • Perform homogeneous screening assays or measure activity secondarily after cell lysis
  • Also compatiible with prepared lysates from other species such as bacteria

Product Details:

Linearity of β-galactosidase assay from cells growing in media in a 96-well plate. Linearity of β-galactosidase assay from cells growing in media in a 96-well plate. Yeast strain BRS1002 carrying plasmid pYX122-β-Gal was grown to an OD600 of 1.0, then 100μL of cells were transferred to a 96-well plate. At time 0, 100μL of a 1:1 mixture of lysis reagent and 2X β-Gal assay buffer were added to each well and absorbance at 420nm was determined. Specific activity for this sample is 160 U (unit = OD405 x 1000/time/OD660).

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.

Related Products:

Mammalian β-Galactosidase Assay Kit
Y-PER Yeast Protein Extraction Reagent


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
75768 Yeast beta-Galactosidase Assay Kit
Formulation: Multi-component kit
Sufficient For: 250 colonies or 250 culture samples (100µL each)

Kit Contents:
Y-PER Yeast Protein Extraction Reagent, 25mL
2X beta-Galactosidase Assay Buffer, 25mL
1M Sodium Carbonate Stop Solution, 25mL
75mL kit Product Instructions for product #75768 Yeast beta-Galactosidase Assay Kit MSDS for product #75768 Yeast beta-Galactosidase Assay Kit Certificate of Analysis for product #75768 Yeast beta-Galactosidase Assay Kit $161.00

Add to cart
75768B Yeast beta-Galactosidase Assay Kit
Formulation: Multi-component kit

Kit Contents:
Y-PER Yeast Protein Extraction Reagent
2X beta-Galactosidase Assay Buffer
1M Sodium Carbonate Stop Solution
Custom MSDS for product #75768B Yeast beta-Galactosidase Assay Kit

Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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