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Updated: Mar 3, 2026
GST-His purification: A Two-step Affinity Purification Protocol Yielding Full-length Purified Proteins
Published on: October 29, 2013
Expression and purification of p70ΔCT104 S6 K, a 72kDa c-terminal truncated p70S6 kinase-GST fusion protein in
Younis Mohammad Hazari1, Irfana Reshi1, Mudasir Habib1
1Department of Biotechnology, University of Kashmir, Srinagar, 190006 Jammu and Kashmir, India.
Abstract:
The p70ΔCT104 S6K is a 421 amino acid residue long truncated form of p70S6 kinase, with 104 amino acids residues cleaved from the carboxyl terminal end of the original protein. The p70ΔCT104 S6K was cloned in E. coli DH5α and successfully expressed in E. coli BL21 (DE3) strain. Western blot with rabbit polyclonal anti-GST antibody was used to follow the protein during expression and purification. The protein purification was achieved by affinity chromatography using Glutathione resin-agarose beads, followed by chromatography on a spin concentration column. The purified protein was confirmed by rabbit polyclonal anti-p70S6 kinase antibody. MALDI/MS Peptide mass fingerprinting confirmed identity of the expressed product.
