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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species Sulfolobus tokodaii
Locus tag STS178
Products conserved hypothetical protein
Plasmid ID 0293
Vector  
Purification Lot STS178_2
Purified Date 2005.6.14-7.21
Protein conc. (mg/ml)  
Total volume (ml)  
Total protein (mg)  

Culture Lot:

date host cell scale label weight of bacteira (g)
        54

Cell Lysis and Heat Treatment:

54 g of E. coli.
180 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
Sonication, output 8, duty 50/2 min , 4 times
Heat 10 min, 75 deg C.
Centrifuge 18,000 g, 60min.
 
 
 
 

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,1.5M AS 1.5M AS 0M AS 6 1.18M AS
2 Resource S 6 ml 50mM MES(pH6.0),5mM B-ME 0M NaCl 2M NaCl 3 0.1M NaCl
3 HiPrep 26/10 Desalting 53ml 20mM CAPS(pH9.0),5mM B-ME,150mM NaCl     1  
4            
5            
6            
7            
8            
9            

Comment:

Tended to precipitate in particular at pH 7.5.

2011.10.20

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