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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species Sulfolobus tokodaii
Locus tag ST1241
Products aspartate kinase
Plasmid ID 0071
Vector  
Purification Lot ST1241_3a
Purified Date 2004.10.25
Protein conc. (mg/ml) 6.68
Total volume (ml) 3.7
Total protein (mg) 24.7

Culture Lot:

date host cell scale label weight of bacteira (g)
        49.0

Cell Lysis and Heat Treatment:

25 g of E. coli cells
140 ml of Ultra Sonication with 20mM Tris-HCl + 50mM NaCl (pH8.0)
Sonication, output 50, duty 8/4 min, 10 times
Heat Treatment at 75 deg C for 15 min
Ultra Cengrifuge at 100,000G for 1 hr
 
 
 
 

Purification Procedure:

Step Column Buffer Gradient 1 Gradient 2 FlowRate (ml/min) Eluted
1 Resource Q 6 mL 20 mM Tris-HCl (pH8.0), 5 mM b-ME 0M NaCl 2M NaCl 6 0 mM NaCl
2 Bio-Scale CHT10I 10 mM KPi (pH7.0), 5 mM b-ME 10mM KPi 500mM KPi 2 157 mM KPi
3 HiLoad 16/60 Superdex 75 20 mM Tris-HCl (pH8.0), 5 mM b-ME, 150 mM NaCl     1  
4 HiPrep 26/10 Desalting 53ml 20 mM Tris-HCl (pH8.0), 5 mM b-ME        
5            
6            
7            
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9            

Comment:

 

2011.10.20

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