2616

http://www.excellgen.net/eln/images/4/4a/2616_2621_digestion.JPG

expression screening

9/27/19

Lane	CLone ID	Name	Induction	Fraction	Expression
1	2615	pCold-MBP-(tev)-GST-(3c)-Ac-LysargiNase	25 oC , 4h	soluble
2	2615		37 oC 4 h	soluble	Y
3	2616	pET28b-hFth-CYFRA 21-1-avitag-6H	25 oC , 4h	soluble	Y
4	2616		37 oC 4 h	soluble	Y
5	2621	pET29a-NSP4-iFlp	25 oC , 4h	soluble
6	2621		37 oC 4 h	soluble
7	2615	pCold-MBP-(tev)-GST-(3c)-Ac-LysargiNase	25 oC , 4h	IB	Y
8	2615		37 oC 4 h	IB	Y
9	2616	pET28b-hFth-CYFRA 21-1-avitag-6H	25 oC , 4h	IB	Y
10	2616		37 oC 4 h	IB	Y
11	2621	pET29a-NSP4-iFlp	25 oC , 4h	IB
M		1250, 130, 100, 70, 55, 35, 25, 15, 10

Lane	ID	Name	Induction	Sample	Expression
1	2555		5052 o/n 37 oCm 1 mM 3 hours 37 oC
2	2556		5052 o/n 37 oCm 1 mM 3 hours 37 oC
3	2557		5052 o/n 37 oCm 1 mM 3 hours 37 oC
4	2558		5052 o/n 37 oCm 1 mM 3 hours 37 oC
5	2559		5052 o/n 37 oCm 1 mM 3 hours 37 oC
6	2560		5052 o/n 37 oCm 1 mM 3 hours 37 oC
7	2561		5052 o/n 37 oCm 1 mM 3 hours 37 oC
8	1562		5052 o/n 37 oCm 1 mM 3 hours 37 oC
9	2563		5052 o/n 37 oCm 1 mM 3 hours 37 oC
10	2564		5052 o/n 37 oCm 1 mM 3 hours 37 oC, cell lysed
11	971		5052 o/n 37 oCm 1 mM 3 hours 37 oC, cell lysed
M
12	2621	pET29a-NSP4-iFlp, 9.76 / 52.8	5052 o/n 37 oCm 1 mM 3 hours 18 oC	S
13	2615	pCold-MBP-(tev)-GST-(3c)-Ac-LysargiNase, 5.23 / 110.7	5052 o/n 37 oCm 1 mM 3 hours 18 oC	S
14	2615	pCold-MBP-(tev)-GST-(3c)-Ac-LysargiNase	5052 o/n 37 oCm 1 mM 3 hours 18 oC	IB

Purification

10/4/19

Expression, LB+ 5052, induce at 18 oC o/n, 2 L

10 ml Ni column.

Wash

Elute Low salt Buffer B-->10 ml Q FF

Elute Q FF with 0--> 1.5 M NaCl.

FLPC, 

http://www.excellgen.net/eln/images/a/af/2616_purification.jpg

Lane	Sample
1	2616 Ni  frac 3
2	2616 Ni  frac 4
3	2616 Ni  frac 5
4	2616 Ni  frac 6
5	2616 Ni  frac 12
6	2616 Q Frac 13
7	2616 Q Frac 14
8	2616 Q Frac 15
M	Page Ruler Plus
9	2624 S
10	2624 secrection
11	?