Cre 298 375 491 1010 1708 1823 2130 2174 375s 1708s 1823s

Protein Expression

2014/7/24
SHANYI

Lane	Name	Protein Name	Clone ID	Host	Induction	Purification	pI/Mw	express	sample
1	pTrix-HTNcre		298	5 ml, Nico21(DE3)plysN	0.4mM IPTG  RT O/N	Ni beads
2	pET28b-HTNcre		375	5 ml, Nico21(DE3)plysN	0.4 mM IPTG RT O/N	Ni beads
3	pET30b-HTNcre		491	5 ml, Nico21(DE3)plysN	0.4mM IPTG RT O/N	Ni beads
4	pBad24-HTNcre		1010	5 ml, Nico21(DE3)plysN	0.2% Arabinose	Ni beads
5	pT7A3-HTNcre		1708	5 ml, Nico21(DE3)plysN	0.4 mM IPTG RT O/N	Ni beads
6	pCold-HTNcre		1823	5 ml, Nico21(DE3)plysN	0.4mM IPTG RT O/N	Ni beads
7	pT7A3-6H-NLS-cre		2130	5 ml, Nico21(DE3)plysN	0.2mM IPTG RT O/N	Ni beads
8	pCold-R9-HA-cre		2174	5 ml, Nico21(DE3)plysN	0.2mM IPTG RT O/N	Ni beads
9	PageRuler Plus Prestained protein ladder
10	pET28b-HTNcre		375	5 ml, Nico21(DE3)plysN	0.4mM IPTG RT O/N	Ni beads
11	pT7A3-HTNcre		1708	5 ml, Nico21(DE3)plysN	0.2mM IPTG RT O/N	Ni beads
12	pCold-HTNcre		1823	5 ml, Nico21(DE3)plysN	0.2mM IPTG RT O/N	Ni beads

other Cre clones

ID	Name	Desc
287	pET8XHis-Cre	NLS-Cre, not HTNCre, no Tat
135	pQE-80L-HTNCre	HTNCre with T5 Promoter
136	pL6H-Tat-Cre	Tat-Cre with pL promoter
1352	pET28b-Cre	Cre (without Tat and NLS)
1312	pET-6H-MPGa-Cre	6H-MPGa-Cre
1311	pET-6H-VPMLK-TAT-NLS-Cre	6H-VPMLK-TAT-NLS-Cre
1310	pET-6H-VPMLK-NLS-Cre	6H-VPMLK-NLS-Cre
1309	pET-6H-KLPVM-NLS-Cre	6H-KLPVM-NLS-Cre
1308	pET-6H-Penetratin-Cre	6H-Penetratin-Cre
1307	pET-6H-VPTLK-NLS-Cre	6H-VPTLK-NLS-Cre
1978	pJ912-HTNCre	yeast
1899	pPinkalpha-HC-HTNCre	yeast
2174	pCold-R9-HA-Cre
2130	pT7A3-6H-NLS-Cre
2074	pET28b-Penetratin-Cre

 

 
 
 
 
 

 

 

Cre Protein Expression

Date:9/20/2018

yijia

Lane	Name	Protein Name	Clone ID	Host	Induction	Modify	Purification	pI/MW	single colony from(Box location)	Sample
M1		PM2500 ExcelBand 3-color Regular Range Protein Marker; in MES buffer;180kD,140kD,100kD,75kD-Red,60kD,45kD,35kD,25kD-Green,15kD,10kD
1	pCold-HTNCre		1823	Nico21	1mM IPTG,RT,o/n	50X 5020	Ni beads		from stock not colony
2	pT7A3-HTNCre		1708	Nico21	1mM IPTG,RT,o/n	50X 5020	Ni beads	10.30 / 42.8	Box 1 F8
3	pT7A3-HTNCre		1708	Nico21	1mM IPTG,RT,o/n	50X 5020	Ni beads	10.30 / 42.8	Box 2 I9
4	pT7A3-HTNCre		1708	Nico21	1mM IPTG,RT,o/n	50X 5020	Ni beads	10.30 / 42.8	Box 14 G6
5	pT7A3-HTNCre		1708	Nico21	1mM IPTG,RT,o/n	50X 5020	Ni beads	10.30 / 42.8	Box 14 H6
6	pT7A3-HTNCre		1708	Nico21	1mM IPTG,RT,o/n	50X 5020	Ni beads	10.30 / 42.8	Box 18 A1
M2	SeeBlue	SeeBlue Protein 4, 6, 14, 17(red), 28, 38, 49, 62, 97(yellow), 191
7		Flpo
8	Negative control	Expi cells					Ni beads
9	positive control	Cre	1708	Nico21	1mM IPTG,RT,o/n		Ni beads
M2	SeeBlue	SeeBlue Protein 4, 6, 14, 17(red), 28, 38, 49, 62, 97(yellow), 191

Cre protein purification QC

Yijia

Lane	where it is from	volumn	Buffer	Note
M	SeeBlue Protein 4, 6, 14, 17(red), 28, 38, 49, 62, 97(yellow), 191	5ul
1	Original lysed fluid	30ul	4XNuPAGE buffer(b-me added)
2	Ni flowthrough	30ul	4XNuPAGE buffer(b-me added)
3	buffer A washing (no immedizole)	30ul	4XNuPAGE buffer(b-me added)
4	buffer A washing(10mM immedizole added)	30ul	4XNuPAGE buffer(b-me added)
5	Elution from Ni	30ul	4XNuPAGE buffer(b-me added)
6	Q flowthrough	30ul	4XNuPAGE buffer(b-me added)
7	SP flowthrough	30ul	4XNuPAGE buffer(b-me added)
8	from SP washing	30ul	4XNuPAGE buffer(b-me added)
9	Elution from SP	30ul	4XNuPAGE buffer(b-me added)
10	BL21 Star induced 1mM IPTG--clone ID 1010(antibiotics A/C)	30ul	2X Sample Buffer
11	Nico21  INduced 1mM IPTG--clone ID 1010(antibiotics A/C)	30ul	2X Sample Buffer