Prev.
Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species Aeropyrum pernix
Locus tag APE0033
Products phosphoenolpyruvate carboxykinase (ATP), pckA
Plasmid ID R4231
Vector  
Purification Lot APE0033
Purified Date 2006.2.16-
Protein conc. (mg/ml)  
Total volume (ml)  
Total protein (mg)  

Culture Lot:

date host cell scale label weight of bacteira (g)
        25.0

Cell Lysis and Heat Treatment:

25 g of E. coli.
↓
100 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 150mM NaCl.
↓
Sonication, output 8, duty 50/2 min, 5 times.
↓
Heat 15min, 75 deg C.
↓
Centrifuge at 100,000 rcf, 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,1.35M AS 1.35M AS 0M AS 5  
2 Resource S 6 ml 50mM MES(pH6.0),5mM B-ME 0M NaCl 0.4M NaCl 4 0.06M NaCl
3            
4            
5            
6            
7            
8            
9            

Comment:

Purified products were not target protein.

2011.10.20

- kuramitsuxxxx_html.pl