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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species Sulfolobus tokodaii
Locus tag ST1131
Products hypothetical protein
Plasmid ID 721
Vector  
Purification Lot ST1131
Purified Date 2006.7.14
Protein conc. (mg/ml) 10.26
Total volume (ml) 1.25
Total protein (mg) 12.75

Culture Lot:

date host cell scale label weight of bacteira (g)
        11.0

Cell Lysis and Heat Treatment:

11.0 g of E. coli.
60ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
Sonication, output 8, duty 50/3min 5 times.
Heat 10 min, 75 deg C.
Centrifuge at 100,000 rcf for 60 min.
70 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,1.0M AS 1.0M AS 1.0M AS 6 between wash and 820mM AS
2 Resource Q 6mL 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 2M NaCl 4 170mM NaCl
3 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 84min
4            
5            
6            
7            
8            
9            

Comment:

Products in the main band was confirmed by Trypsin-PMF.

2011.10.20

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