Prev.
Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST0600
Products  
Plasmid ID 0032
Vector  
Purification Lot ST0600
Purified Date 2004.10.14
Protein conc. (mg/ml) 11.44
Total volume (ml) 3.75
Total protein (mg) 42.9

Culture Lot:

date host cell scale label weight of bacteira (g)
        26.7

Cell Lysis and Heat Treatment:

26.7 g of E. coli.
105 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
Sonication, output 8/duty 50, 2 min, 4 times.
Heat 15min, 75 deg C.
Centrifuge at 100,000 rcf for 60min.
88 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,1.35M AS 1.35M AS 0M AS 6 930mM AS
2 BioAssist S 10X10 50mM MES(pH6.0),5mM B-ME 0M NaCl 1M NaCl 2 450mM NaCl
3 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     0.5 72ml
4            
5            
6            
7            
8            
9            

Comment:

Products were confirmed by Trypsin-Mass MSCOT.

2011.10.20

- kuramitsuxxxx_html.pl