Plasmid and Protein Purification Database -Purification-

Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species
Locus tag	ST0182
Products
Plasmid ID	0402
Vector
Purification Lot	ST0182_1a
Purified Date	2006.3.8
Protein conc. (mg/ml)	4.30
Total volume (ml)	0.42
Total protein (mg)	1.8

Culture Lot:

date	host cell	scale	label	weight of bacteira (g)
				10.8

Cell Lysis and Heat Treatment:

10.8 g of E. coli cell.

↓

70 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.

↓

Purification Procedure:

Step	Column	Buffer	Gradient 1	Gradient 2	FlowRate (ml/min)	Eluted
1	Resource Iso 6mL	20mM Tris-HCl(pH8.0),5mM B-ME,1.5M AS	1.5M AS	0M AS	3	0.9M AS
2	Resource S 6mL	50mM MES(pH6.0),5mM B-ME	0M NaCl	300mM NaCl	4	120mM NaCl
3	HiLoad 16/60 Superdex 75	20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl			1	68mL/120mL
4
5
6
7
8
9

Comment:

Solution was divided into three, a, b, and c, at the end of Purification 2. Products were confirmed by Trp-Mass-Mascot. A product of 44 kDa also identified as the target protein.

2011.10.20

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