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 Sridharan 1, Tripthi Battapadi 1, Lata Balakrishnan 1
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 Battapadi 1, Madhumita Sridharan 1, Degang Liu 2, John Turchi 3, Lata Balakrishnan 1
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 (KD) of the interaction.
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