Paul.xu: Created page with "Date: 2014/2/20
Cells: BL21-AI, 5 ml, induce with 0.15 mM o/n at 18 oC. (cell initially all lysed, only a few survived, then growed to OD600 ~=1 at 37 oC, cool down to ~20 o..."

2014-02-23T22:59:01Z

Created page with "Date: 2014/2/20 Cells: BL21-AI, 5 ml, induce with 0.15 mM o/n at 18 oC. (cell initially all lysed, only a few survived, then growed to OD600 ~=1 at 37 oC, cool down to ~20 o..."

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Date: 2014/2/20 

Cells: BL21-AI, 5 ml, induce with 0.15 mM o/n at 18 oC. (cell initially all lysed, only a few survived, then growed to OD600 ~=1 at 37 oC, cool down to ~20 oC, add IPTG to 0.15 mM). 

Purification: 50 ul Ni beads 

S: cell lysate supernatant, beads elute with 25 ul 0.3 M Imidazole, first elute 

S2: beads elute with 25 ul 0.3 M Imidazole, 2nd elute 

P: cell lysate pellet, add 8 M urea to dissolve the pellet, add 50 ul Ni beas, beads elute with 25 ul 0.3 M Imidazole, first elute

P2: beads elute with 25 ul 0.3 M Imidazole, 2nd elute 

 

[[Image:2096_2097_expression_screening.jpg]] 

 

Conclusion: Yeast version of Flp is better than E coli optimized high CAI version 

[[Category:Protein_Purification]]

2096 2097 - Revision history

Paul.xu: Created page with "Date: 2014/2/20 Cells: BL21-AI, 5 ml, induce with 0.15 mM o/n at 18 oC. (cell initially all lysed, only a few survived, then growed to OD600 ~=1 at 37 oC, cool down to ~20 o..."

Paul.xu: Created page with "Date: 2014/2/20
Cells: BL21-AI, 5 ml, induce with 0.15 mM o/n at 18 oC. (cell initially all lysed, only a few survived, then growed to OD600 ~=1 at 37 oC, cool down to ~20 o..."