Lab _4 - Beta-Gal (Precipitation)

Lab _4 - Beta-Gal (Precipitation) - Purification &...

Info iconThis preview shows pages 1–6. Sign up to view the full content.

View Full Document Right Arrow Icon
Next three lab sessions: 1 st Lab : salting out of β-gal and determine ideal β-gal concentration range to work with. nd                          of  E. coli    β - galactosidase ( β -gal)
Background image of page 1

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Objectives: to develop a familiarity with serial dilution procedure. to determine the ideal concentration of β-gal for the measurements of enzyme activity and protein concentration (Bradford assay). to separate β-gal from other cellular components by salting-out .                        of  E. coli    β -galactosidase   (session 1)
Background image of page 2
β -gal is encoded by the Lac Z gene of the  lactose (lac) operon β -Galactosidase
Background image of page 3

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
o-NPG Galactose + o-NP (o-Nitrophenyl- β , d-galactopyranoside) (o-Nitrophenol) (colorless) (bright yellow, 420 nm) β -gal β -gal is an essential enzyme in the  metabolism of lactose β -gal also hydrolyzes a lactose substrate analog, o-NPG. The rate of o-NP production from o-NPG is directly proportional to the amount of β -gal in the sample.
Background image of page 4
The cell extract is obtained by the following procedure: 1. E. coli , strain ML-308, was grown overnight in liquid broth.
Background image of page 5

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Image of page 6
This is the end of the preview. Sign up to access the rest of the document.

Page1 / 13

Lab _4 - Beta-Gal (Precipitation) - Purification &...

This preview shows document pages 1 - 6. Sign up to view the full document.

View Full Document Right Arrow Icon
Ask a homework question - tutors are online