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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST1495
Products  
Plasmid ID 0093
Vector  
Purification Lot ST1495
Purified Date 2004.7-
Protein conc. (mg/ml) 11.6
Total volume (ml) 2.0
Total protein (mg) 23.2

Culture Lot:

date host cell scale label weight of bacteira (g)
        31.8

Cell Lysis and Heat Treatment:

31.8 g of E. coli.
140 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
Sonication, output 8/duty 50, 1.5 min, 7 times.
Heat 10min, 70 deg C.
Centrifuge at 100,000 rcf for 60min.
110 ml of supernatant.
 
 
 

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,1.05M AS 1.05M AS 0M AS 6 0.9M AS
2 Resource Q 6 ml 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 2M NaCl 4 290mM NaCl
3 Bio-Scale CHT10I 10mM KPi(pH7.0),5mM B-ME 10mM KPi 500mM KPi 2 68.8mM KPi
4 HiPrep 26/10 Desalting 53ml 20mM Tris-HCl(pH8.0),5mM B-ME        
5            
6            
7            
8            
9            

Comment:

Products were confirmed by MS.

2011.10.20

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