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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species Sulfolobus tokodaii
Locus tag ST1531
Products pyruvate ferredoxin oxidoreductase, gamma subunit
Plasmid ID R0232
Vector  
Purification Lot ST1531
Purified Date 2006.6.19-6.28
Protein conc. (mg/ml) 13.26
Total volume (ml) 0.75
Total protein (mg) 10.4

Culture Lot:

date host cell scale label weight of bacteira (g)
        81.5

Cell Lysis and Heat Treatment:

Combined lysates of 40.5 g and 41.0 g E. coli.
200 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
Sonication, output 8, duty 50/2min, 5 times.
Heat 15 min 75 deg C.
Centrifuge at 100,000 rcf for 50 min.
200 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 5 1.3M AS
2 Resource Q 6 ml 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 0.6M NaCl 5 180mM NaCl
3 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 83ml/120ml
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Comment:

Products was confirmed by Trp-MS.

2011.10.20

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