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

Madhumita Sridharan; Tripthi Battapadi; Lata Balakrishnan

doi:10.3791/66529

Rivista

Biochimica

Optimizing FPLC for Efficient Purification of His-Tagged FEN1 Protein

April 26, 2024

Author Spotlight: Optimizing Affinity Chromatography for His-Tagged FEN1 Protein

01:19 min

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Affinity Column Preparation and Packing for Fast Protein Liquid Chromatography (FPLC)

01:42 min

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Priming the Sample Loop of the Fast Protein Liquid Chromatography (FPLC) System for the Purification of His-Tagged FEN1 Protein

01:26 min

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Purification of His-Tagged FEN1 Protein Using the FPLC System and Its Analysis Using FPLC-Linked Software

02:58 min

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Madhumita Sridharan¹, Tripthi Battapadi¹, Lata Balakrishnan¹

¹Department of Biology,Indiana University Indianapolis

Summary

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.