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Plasmid & Protein Purification Database -Purification-

Species Sulfolobus tokodaii
Locus tag ST0971
Products hypothetical sulfide dehydrogenase [flavocytochrome c] flavoprotein subunit
Plasmid ID 0495
Vector  
Purification Lot ST0971_1
Purified Date 2005.4.28
Protein conc. (mg/ml) 10.6
Total volume (ml) 0.92
Total protein (mg) 9.75

Culture Lot:

date host cell scale label weight of bacteira (g)
        20.0

Cell Lysis and Heat Treatment:

20.0 g of E. coli.
100ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
Sonication, output 8, duty 50/2 min, 5 times.
Heat 15 min, 80 deg C.
Centrifuge at 100,000 rcf, 50 min.
90 ml of supernatant.
 
 
 

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 3 between 620mM AS and 250mM AS
2 BioAssist S 10X10 50mM MES(pH6.0),5mM B-ME 0M NaCl 500mM NaCl 1.5 50mM NaCl
3 HiLoad 16/60 Superdex 200 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 73ml
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Comment:

 

2011.10.20

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