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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST2055
Products  
Plasmid ID R254b
Vector  
Purification Lot ST2055_6
Purified Date 2005.6.28-7.5
Protein conc. (mg/ml) 10.7
Total volume (ml) 0.73
Total protein (mg) 7.8

Culture Lot:

date host cell scale label weight of bacteira (g)
        40.0

Cell Lysis and Heat Treatment:

40.0 g of E. coli.
130 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
Sonication, output 8, duty 50/2 min, 5 times.
Heat 10 min, 75 deg C.
Centrifuge at 100,000 rcf, 60 min.
120 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,1.8M AS 1.8M AS 0M AS 6 1.7M AS
2 Resource S 6 ml 50mM MES(pH6.0),5mM B-ME 0M NaCl 2M NaCl 4 69mM NaCl
3 Bio-Scale CHT5I 10mM KPi(pH7.0),5mM B-ME 10mM KPi 500mM KPi 2 83mM KPi
4 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME     1  
5            
6            
7            
8            
9            

Comment:

 

2011.10.20

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