8.29
Pick Clon
8.30
Pilot Expression of Protein
“IPTG+ 4h”“IPTG- 4h”
After the rocker culture is 4 H, add IPTG(final concentration is 0.5 mm) and then the rocker culture is 4 hours. Each of the two tubes absorbs 200 μL of bacterial fluid, and in the frozen centrifuge, it is 4 °C and 12000 rpm centrifuge. 10 minutes, go to the clear, It is resuspended with 100 μL 1x SDS Loading buffer, then heated in a water bath pot at 100 °C for 10 minutes and stored in a -20 °C refrigerator.
8.31
Protein extraction
CA2L203K
CA2WT
CA2L203K
9.01
Purification of Protein
Prior to purification
Purified
9.03
Western blot identification of proteins
CA2L203K CA2WT