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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species Sulfolobus tokodaii
Locus tag ST0075
Products hypothetical NAD-dependent alcohol dehydrogenase
Plasmid ID 0397
Vector  
Purification Lot ST0075
Purified Date 2006.3.6
Protein conc. (mg/ml) 9.43
Total volume (ml) 1.02
Total protein (mg) 9.62

Culture Lot:

date host cell scale label weight of bacteira (g)
        8.4

Cell Lysis and Heat Treatment:

8.4 g of E. coli cell.
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70.3 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
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Sonication, output 8, duty 50/3min, 10 times.
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Heat 15min, 75 deg C.
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Centrifuge at 100,000 rcf, 50 min.
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Purification Procedure:

Step Column Buffer Gradient 1 Gradient 2 FlowRate (ml/min) Eluted
1 Resource Phe 6mL 20mM Tris-HCl(pH8.0),5mM B-ME,1.35M AS 1.35M AS 0M AS 4 1.18M AS
2 Resource S 6mL 50mM MES(pH6.0),5mM B-ME 0M NaCl 500mM NaCl 4 150mM NaCl
3 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 67min
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Comment:

 

2011.10.20

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