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Plasmid & Protein Purification Database -Purification-

Species Sulfolobus tokodaii
Locus tag ST0059
Products hypothetical selenium-binding protein
Plasmid ID 0701
Vector  
Purification Lot ST0059_1
Purified Date 2006.8.17
Protein conc. (mg/ml) 46.40
Total volume (ml) 1.31
Total protein (mg) 60.78

Culture Lot:

date host cell scale label weight of bacteira (g)
        25.6

Cell Lysis and Heat Treatment:

25.6 g of E. coli cell.
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140ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
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Purification Procedure:

Step Column Buffer Gradient 1 Gradient 2 FlowRate (ml/min) Eluted
1 Resource Iso 6mL 20 mM Tris-HCl (pH8.0), 5 mM b-ME, 1.5 M AS 1.5 M AS 0M AS 5 1.0 M AS
2 Resource Q 6mL 20 mM Tris-HCl (pH8.0), 5 mM b-ME 0M NaCl 600mM NaCl 4 120mM NaCl
3 HiLoad 16/60 Superdex 200 20 mM Tris-HCl (Ph 8.0), 5 mM b-ME, 150 mM NaCl     1 75mL/120mL
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Comment:

Products without heat treatment were run faster on a SDS-PAGE gell. Products was confirmed by MS after trypsinization.

2011.10.20

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