Indiana University Indianapolis

2 ARTICLES PUBLISHED IN JoVE

Biochemistry

Affinity Purification of a 6X-His-Tagged Protein using a Fast Protein Liquid Chromatography System

Madhumita Sridharan¹, Tripthi Battapadi¹, Lata Balakrishnan¹

¹Department of Biology, Indiana University Indianapolis

This article provides a procedure for the affinity purification of a human recombinant protein, flap endonuclease 1 (FEN1), which has been labeled with a 6X-histidine tag. The protocol involves the utilization of two distinct immobilized metal ion columns for the purification of the tagged protein.

Biochemistry

Analyzing DNA-Protein Interactions with Streptavidin-Based Biolayer Interferometry

Tripthi Battapadi¹, Madhumita Sridharan¹, Degang Liu², John Turchi³, Lata Balakrishnan¹

¹Department of Biology, Indiana University Indianapolis, ²Sartorius Corporation, ³Department of Medicine, Indiana University School of Medicine

This article describes a protocol for studying DNA-protein interactions using a streptavidin-based biolayer interferometry (BLI) system. It outlines the essential steps and considerations for utilizing either basic or advanced binding kinetics to determine the equilibrium binding affinity (K_D) of the interaction.