Protein Purification Techniques

Question 1

Explain E.coli cell lysis (step 1)

Answer 1

resuspend pelleted cells in a Lysis buffer with pH=7.4, 10mM imidazole to reduce non-specific IMAC binding, DNAsel to reduce viscosity and more efficiently filter the solution, protease inhibitor, lysozyme.

Then separate the lysate with a centrifuge and filter the supernatant

Question 2

Explain IMAC

Answer 2

IMAC - Immobilized Metal Chelate Affinity Chromatography.

filter supernatant onto a Nickel resin-column and elute it.