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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species Sulfolobus tokodaii
Locus tag ST1506
Products conserved hypothetical protein
Plasmid ID R99
Vector  
Purification Lot ST1506_1
Purified Date 2006.3.28-4.6
Protein conc. (mg/ml) 0.93
Total volume (ml) 0.84
Total protein (mg) 0.78

Culture Lot:

date host cell scale label weight of bacteira (g)
        11.2

Cell Lysis and Heat Treatment:

11.2 g of E. coli.
72.5 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
Sonication, output 8, duty 50/3min, 10 times.
Heat 15 min, 75 deg C.
Centrifuge at 100,000 rcf for 60 min.
60 ml of supernatant.
 
 
 

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,2.0M AS 2.0M AS 0M AS 4 1.65M AS
2 Resource Q 6 ml 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 1M NaCl 4 0.42M NaCl
3 Bio-Scale CHT2I 10mM KPi(pH7.0),5mM B-ME 10mM KPi 500mM KPi 1 238.5mM KPi
4 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME,0.5M NaCl     1 55min
5            
6            
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9            

Comment:

Products was confirmed by MS.

2011.10.20

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