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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species Sulfolobus tokodaii
Locus tag ST1819
Products precorrin-8W decarboxylase
Plasmid ID R469
Vector  
Purification Lot ST1819
Purified Date 2006.9.12-9.19
Protein conc. (mg/ml) 20.6
Total volume (ml) 1.24
Total protein (mg) 25.5

Culture Lot:

date host cell scale label weight of bacteira (g)
        20.5

Cell Lysis and Heat Treatment:

20.5 g of E. coli.
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100 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
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Sonication, output 8, duty 50/4 min, 5 times.
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Heat 10 min, 75 deg C.
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Centrifuge at 100,000 rcf, 60 min.
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94 ml of supernatant.
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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 6 1.79M AS
2 Resource Q 6 ml 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 1.30M NaCl 4 0.11M NaCl
3 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 71min
4            
5            
6            
7            
8            
9            

Comment:

Products in the main band was confirmed by Trypsin-PMF.

2011.10.20

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