TLS Online TPP Program

#Id: 2843

#Unit 13. Methods in Biology

Recovery of proteins using (6His-Tag) and (Ni-NTA) chromatography columns.

TLS Online TPP Program

#Id: 2844

#Unit 13. Methods in Biology

The glucose uniporter GLUT1 can be purified from the solubilized mixture by antibody affinity chromatography on a column containing either a GLUT1-specific monoclonal antibody or an antibody specific for the epitope tag, then incorporated into liposomes made of pure phospholipids.

TLS Online TPP Program

#Id: 2845

#Unit 13. Methods in Biology

Affinity chromatography fraction can be analyzed by two ways

Absorption or UV visible spectrophotometry which analyzes the concentration of protein in the fraction by using Beer lamberts law.

Polyacrylamide gel electrophoresis to fractionize the protein sample on the basis of their molecular weight.

TLS Online TPP Program

#Id: 2846

#Unit 13. Methods in Biology

Sample injection; due to the high protein concentration, the absorbance reaches the maximum recordable value; most protein passes through the column, unbound.

Contaminating proteins eluting by way of a salt gradient.

TLS Online TPP Program

#Id: 2847

#Unit 13. Methods in Biology

In Size exclusion chromatography (SEC) molecules dissolved in the matrix or stationary phase or high retention time or small molecular weight elute last.

TLS Online TPP Program

#Id: 2848

#Unit 13. Methods in Biology

SEC chromatographic technique for the separation of molecules on the basis of their molecular size and shape exploits the molecular sieve properties of a variety of porous materials.