IFN-α or -β activates IFNAR to recruit and activate specific JAKs (JAK1 and Tyk2), which then activate specific STATs induce expression of three genes, Protein kinase R (PKR), which inhibits viral (and cellular) protein synthesis by inhibiting the elongation factor eIF2α; 2’,5’-oligoadenlyate A synthetase, which activates the ribonuclease RNase L that degrades viral mRNA; and Mx proteins, which inhibit both the transcription of viral genes into mRNAs and the assembly of virus particles.

TLS Online TPP Program

#Id: 2842

#Unit 13. Methods in Biology

GLUT separation is also done with the help of affinity chromatography, normally glut protein are cloned into the vector and then the help of affinity chromatography

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.