Cre pTriEx-HTNC-purification: Difference between revisions
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Latest revision as of 16:52, 7 February 2012
Date: 2011-11-14 to 2011-11-17
Notebook: Ruili Su-- Protein purification
Page:002857 to 002859
Title: pTriEx-HTNC (Clone ID298) protein expression and purification.
-11-16-2011.jpg)
PTriEx-HTNC: use plasmid (Clone ID298) to transform with Rosetta BL21 (DE3) pLysS competent cells and culture it on plate containing Chl/Car. Grow 1L at 30oC for O/N, then induce for 6 ~7 hours by IPTG at 30oC. The bacteria growth takes one night and one day.
The sample is just from the washed procedure with washing buffer (50mM imidazole) and still not very pure. So, the imidazole concentration needs to be adjusted before the protein is completely eluted.
_protein----Jan.03_2012.jpg)
Notebook: ruili su 2
Page: 003487
Date: 01/03/2012
PTriEx-HTNC transformed with C41 (DE3)PlysS. After FPLC, runing the SDS-PAGE.
Result
The expression level is good and the purity of Cre protien is good as well.