1708 1823 2116 2133 2115 2108

Expression screening, 4/2/2014, by Shanyi

PageRuler Plus 10, 15, 25(red), 35, 55, 70(red), 100, 130, 250

0.5 mM IPTG induction o/n at RT, 5 ml culture

Lyse cell by freeze-thawing, 3 cycles, using Buffer A, spin--> Supernatant (S)

Add lysis buffer containing 4M urea, 1% SDS in Buffer A to pellet, solubilize and spin-->P

Lane	Clone ID	Name	Competent Cells	Sample	Expression	Note
1	1708	pT7A3-HTNCre	Tuner(DE3)	S
2	1708			P	IB?
3	1823	pCold-HTNCre	Nico(DE3)pLysN	S	Soluble
M		Fermentas	PageRuler Plus
4	1823			P
5	1823	pCold-HTNCre	Tuner(DE3)	S
6	1823			P	IB?
7	2116	pT7A3-Penetratin-Cre	Tuner(DE3)	S	No
8	2116			P
9	2133	pT7A3-ILK	BL21AI-pRare2LysS	S
10	2133			P	low
11	2133	pT7A3-ILK	Tuner(DE3)-pRare2LysS	S	Soluble

Lane	Clone ID	Name	Competent Cells	Sample	Expression	Note
1	2133	pT7A3-ILK	Tuner(DE3)-pRare2LysS	P	some
2	2115	pT7A3-iFLP	Tuner(DE3)	S
3	2115			P	IB?
4	2115		Nico(DE3)pRarepLysS	S
5	2115			P	IB?, low
6	2115		Nico(DE3)pLysN	S		cell death
M	Fermentas					PageRuler Plus
7	2115			P		cell death
8	2115		BL21(DE3)pRareLysS	S
9	2115			P	IB?
10	2108	pCold-Tat-NLS-iFLP-6H	Tuner(DE3)	S
11	2108			P	IB?