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Plasmid & Protein Purification Database -Purification-

Species Sulfolobus tokodaii
Locus tag ST1092
Products hypothetical insertion element protein
Plasmid ID 0197
Vector  
Purification Lot ST1092_b-1
Purified Date 2007.3.1
Protein conc. (mg/ml) 4.54
Total volume (ml) 4.95
Total protein (mg) 22.5

Culture Lot:

date host cell scale label weight of bacteira (g)
        13.2

Cell Lysis and Heat Treatment:

13.2 g of E. coli.
66ml of 20mM Tris-HCl (pH8.0), 50mM NaCl, 5mM B-ME.
Sonication, output 8, duty 50/4min, 10 times.
Heat 10 min, 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,1.75M AS 1.75M AS 0M AS 4 1.3M AS
2 Resource S 6mL 50mM MES(pH6.0),5mM B-ME 0M NaCl 600mM NaCl 4 460mM NaCl
3 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 83min
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Comment:

 

2011.10.20

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