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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species  
Locus tag STS042
Products  
Plasmid ID 0155
Vector  
Purification Lot STS042_4
Purified Date 2005.9.1-9.7
Protein conc. (mg/ml) 10.1
Total volume (ml) 1.69
Total protein (mg) 17.1

Culture Lot:

date host cell scale label weight of bacteira (g)
        20.7

Cell Lysis and Heat Treatment:

20.7 g of E. coli.
102.1 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 150mM NaCl.
Sonication, output 8, duty 5/2 min, 5 times.
Heat 15 min, 75 deg C.
Centrifuge at 100,000 rcf, 50 min.
95 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,2.0M AS 2.0M AS 0M AS 6 from 1.93M AS to 1.58M AS
2 BioAssist Q 10x10 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 0.25M NaCl 2 from 0.02M NaCl to 0.04M NaCl
3 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 66ml
4            
5            
6            
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8            
9            

Comment:

There were two peaks. Products in the main peak assumed that contains multimerized products were recovered.

2011.10.20

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