Cre expression screening

Protein Expression

2014/10/17
SHANYI

Lane	Name	Protein Name	Clone ID	Host	Induction	Purification	pI/Mw	express	note
1	pET28b-HTNCre	cre	375	Nico21(DE3)plysN	0.5 mM IPTG 30oC 4.5h	Ni beads			2g/500ml suspend in 40ml BufferA,take 1ml to do protein expression screening
2	pET28b-HTNCre		375	RossetaplySs	0.5 mM IPTG 10ml 50x5020 RT O/N	Ni beads			3.8g/50ml suspend in 45ml BufferA,take 1ml to do protein expression screening
3	pT7A3-HTNCre		1708	Nico21(DE3)plysN	0.5 mM IPTG 30oC 4.5h	Ni beads			3g/500ml suspend in 45ml BufferA,take 1ml to do protein expression screening
4	pCold-HTNCre		1823	Nico21(DE3)plysN	0.5 mM IPTG RT 5.5h	Ni beads			3g/500ml suspend in 45ml BufferA,take 1ml to do protein expression screening
5	pBad24-HTNCre		1010	Nico21(DE3)plysN	0.2%Arabinose 30oC 6h	Ni beads			7.7g/1.25L suspend in 110ml BufferA,take 1ml to do protein expression screening
6	pTriEx-HTNCre		298	BL21(DE3) pLysS	0.5 mM IPTG 30oC ?h	Ni beads			grow very slowly
7	pET28b-HTNCre		375	Nico21(DE3)plysN	0.5 mM IPTG 37oC 2h	Ni beads			pellet from 8L purification
8	PageRuler Plus Prestained protein ladder
9		HTNCre	1708	Nico21(DE3)plysN					10mM Imidazol wash flowthrough
10			1708	Nico21(DE3)plysN	0.5 mM IPTG 37oC 2h	Ni beads			20mM Imidazol wash flowthrough
11			298	BL21(DE3) pLysS	0.5 mM IPTG 37oC 2h	Ni beads		10mM Imidazol wash flowthrough
12			298	BL21(DE3) pLysS	0.5 mM IPTG 37oC 2h				20mM Imidazol wash flowthrough

10/1/2018

Lane	Name	Protein Name	Clone ID	Host	Induction	Purification	pI/Mw	express	note
1	M
2	Original lysis cell fluid	Cre	1708	NIco21	1mM IPTG,o/n,	Ni beads
3	Ni flow through
4	From Buffer A Wash
5	From immedizole buffer A wash
6	Ni elution
7	Q flowthrough
8	SP Flow through
9	From SP washing buffer wash
10	SP Elution
11	From Nico 21	cre	1010	Nico21		Ni Beads
12	From BL 21	cre	1010	BL21 star		Ni Beads

2/17/19