SlyD removal from Ni-NTA purified protein with nanoCLAMP slyD-A1(Resin)


Overview
Abstract
E. coli SlyD (FKBP-type peptidyl-prolyl cis-trans isomerase) is a common contaminant of 6xHis-tagged proteins purified with Ni-NTA immobilized metal affinity chromatography (IMAC). This application describes the removal of SlyD contaminant from a 6xHis tagged protein by incubation with slyD-A1(Resin). Use of nanoCLAMP slyD-A1(Resin) for removing SlyD protein represents an alternative to expression of 6xHis-tagged proteins in specialized E. coli strains such as LOBSTR, NiCo21(DE3), and NiCo22(DE3).
Type
  • Affinity Chromatography
Target
Scores
Outcome
Successful
Signal
Acceptable
Cross Reactivity
Acceptable
Background
Acceptable
Dynamic Range
Acceptable
Results
Summary
Incubation of the SlyD contaminated protein sample with slyD-A1(Resin) resulted in the depletion of SlyD to below the limits of detection measured by Coomassie staining of approximately 10 ug of target protein separated by SDS-PAGE.
Figure

Image 

SDS PAGE slyD removal from 6xHis tagged protein with nanoCLAMP slyD-A1

Legend 

Depletion of SlyD contaminant from a Ni-NTA purified protein sample. A protein purified by Ni-NTA chromatography (IMAC prep) was incubated with a non-SlyD specific nanoCLAMP(Resin) and with slyD-A1(Resin), allowed to flow through the resin, and analyzed on Coomassie-stained SDS-PAGE.