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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST0238
Products  
Plasmid ID 0152
Vector  
Purification Lot ST0238
Purified Date 2006.2.14
Protein conc. (mg/ml) 37.9
Total volume (ml) 2.78
Total protein (mg) 105.4

Culture Lot:

date host cell scale label weight of bacteira (g)
        12.2

Cell Lysis and Heat Treatment:

12.2 g of E. coli cell.
↓
110.3 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
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Sonication, output 8, duty 50/3min, 10 times.
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Heat 15min, 75 deg C.
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Centrifuge at 100,000 rcf, 60 min.
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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,2M AS 2M AS 0M AS 4 1.69M AS
2 Resource S 6mL 50mM MES(pH6.0),5mM B-ME 0M NaCl 2M NaCl 4 56mM NaCl
3 HiTrap Heparin HP 10mM KPi(pH6.5),5mM B-ME 10mM KPi 500mM KPi 2 Flow through
4 HiLoad 16/60 Superdex 75 50mM MES(pH6.0),5mM B-ME,150mM NaCl     1 70 min
5            
6            
7            
8            
9            

Comment:

A contaminant appeared below the subjected protein in the SDS-PAGE gel was identified as a degradation product of subjected protein.

2011.10.20

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