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This study demonstrates that high levels of a series of recombinant self-assembling peptides can be purified using a simple process for applications as scaffolds in tissue engineering.
Our results show for the first time that human photoreceptors derived from either ES cells or iPS cells can be purified using a combination of photoreceptor-specific GFP vector and fluorescent activated cell sorting.
The resulting ~285 bp product was cloned into the pRSET B vector in order to permit prokaryotic expression of N-terminally His-tagged rGRA5, which could be purified using a nickel resin column.
If a target protein is produced in fusion with a His-tag sequence, it can be purified using a solid support that is covalently modified to displays a heavy metal ion like Ni2+ or Zn2+ on the surface.
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Acetonitrile (MeCN) was purified using a known procedure [39, 40].
PCR products were purified using a PCR Clean-Up Kit (GMbiolab).
The protease fractions were purified using a 2D-Cleanup Kit.
The Pu stock solution was purified using a TEVA column.
The water was purified using a Millipore filtration system.
The product was purified using a PD-10 column and RCP was determined with SEC-HPLC.
DDI water was purified using a Barnstead E-pure (Model D4641) water system.
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