Prev.
Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species  
Locus tag APE0550
Products  
Plasmid ID R4333
Vector  
Purification Lot APE0550_9
Purified Date 2006.5.31-6.12
Protein conc. (mg/ml)  
Total volume (ml)  
Total protein (mg)  

Culture Lot:

date host cell scale label weight of bacteira (g)
        165

Cell Lysis and Heat Treatment:

25.2 g of E. coli.
142.8 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 150mM NaCl.
Sonication, output 8, duty 50/4 min, 10 times.
Heat 15 min, 75 deg C.
Centrifuge at 100,000 rcf for 60 min.
 
 
 
 

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,1.65M AS 1.65M AS 0M AS 5  
2 Resource Q 6 ml 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 0.6M NaCl 4  
3 HiLoad 16/60 Superdex200 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl        
4            
5            
6            
7            
8            
9            

Comment:

 

2011.10.20

- kuramitsuxxxx_html.pl