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New England Biolabs 2020. Separating the Protein of Interest from MBP after TEV Protease Cleavage (NEB #E8201). protocols.io https://dx.doi.org/10.17504/protocols.io.bfebjjanCopy Citation Copied
URL: https://dx.doi.org/DOI:10.17504/protocols.io.bfebjjan
Authors: New England Biolabs
Group: New England Biolabs (NEB)
Summary: Both the MBP-tag and TEV Protease are polyhistidine-tagged for easy removal from the TEV Protease reaction. Loading the digest onto NEBExpress Ni Resin (NEB #S1428) sequesters both the MBP-tag and TEV Protease, thereby isolating the protein of interest in the column flow through. The binding capacity of NEBExpress Ni Resin is ≥ 10 mg/ml. The binding capacity will vary depending on the target protein, binding conditions and the accessibility of the His-tag. It is recommended to estimate the amount of the His-tagged MBP released by TEV Protease cleavage by first analyzing a sample by SDS-PAGE and comparing its band size and intensity to that of the provided MBP6 protein control.
Affiliations: New England Biolabs
External URL: https://www.neb.com/protocols/2020/02/06/separating-the-protein-of-interest-from-mbp-after-tev-protease-cleavage-neb-e8201
Version: 1
Publication Date: 2020
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