Are you sure you want to leave this community? Leaving the community will revoke any permissions you have been granted in this community.
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
Expand AllComing soon.
Source: