Question

Why did we need to measure the absorbance at two wavelengths during the beta-galactosidase lab?

Answer 1

While the absorbance at 420 nm is related to the amount of o-nitrophenol produced, the absorbance at 600 nm is proportional to cell density, which aids in standardizing our estimations of enzyme activity.

Describe absorbance.

The amount of light absorbed by a solution is measured by its absorbance (A), often referred to as optical density (OD). The amount of light that may flow through a solution is called its transmittance.

How is the activity of beta-galactosidase determined?

The colorless ONPG substrate is changed by beta-Galactosidase into galactose and the chromophore o-nitrophenol, which results in a vivid yellow solution. The amount of substrate transformed at 420 nm can be calculated by measuring the solution's beta-galactosidase activity using a spectrophotometer or a microplate reader.

What is measured by the beta-galactosidase assay?

The -Gal Assay Kit gives users the tools they need to swiftly assess the amounts of active beta-galactosidase expressed in cells that have been transfected with plasmids encoding the lacZ gene.

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