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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST1411
Products  
Plasmid ID BL221
Vector  
Purification Lot ST1411
Purified Date 2006.11.11.16
Protein conc. (mg/ml) 33.1
Total volume (ml) 1.48
Total protein (mg) 49.0

Culture Lot:

date host cell scale label weight of bacteira (g)
        26

Cell Lysis and Heat Treatment:

26.0 g of E. coli.
↓
135 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
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Sonication, output 8/duty 50, 4 min, 10 times.
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Heat 15min, 75 deg C.
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Centrifuge at 100,000 rcf for 60min.
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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,1.8M AS 1.8M AS 0M AS 4 1.53M AS
2 Resource Q 6 ml 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 0.6M NaCl 4 Flow through
3 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 73min
4            
5            
6            
7            
8            
9            

Comment:

Products were confirmed by Trp-MS.

2011.10.20

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