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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST0985
Products  
Plasmid ID 0496
Vector  
Purification Lot ST0985
Purified Date 2005.6.7-6.14
Protein conc. (mg/ml) 12.7
Total volume (ml) 2.87
Total protein (mg) 36.4

Culture Lot:

date host cell scale label weight of bacteira (g)
        18.0

Cell Lysis and Heat Treatment:

18.0 g of E. coli.
100 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
Sonication, output 8/duty 50, 1 min, 5 times.
Heat 10min, 75 deg C.
Centrifuge at 100,000 rcf for 60min.
83 ml of supernatant.
 
 
 

Purification Procedure:

Step Column Buffer Gradient 1 Gradient 2 FlowRate (ml/min) Eluted
1 Resource Phe 6mL 20mM Tris-HCl(pH8.0),5mM B-ME,2.0M AS 2.0M AS 0M AS 4 830mM AS
2 Resource Q 6mL 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 200mM NaCl 2 50mM NaCl
3 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 76ml/120ml
4            
5            
6            
7            
8            
9            

Comment:

Products were confirmed by Trypsin-Mascot.

2011.10.20

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