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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species Sulfolobus tokodaii
Locus tag ST1894
Products hypothetical glycine cleavage system H protein
Plasmid ID R0542
Vector  
Purification Lot ST1894
Purified Date 2005.12.19-2006.1.12
Protein conc. (mg/ml) 25.5
Total volume (ml) 2.12
Total protein (mg) 54.06

Culture Lot:

date host cell scale label weight of bacteira (g)
        20.4

Cell Lysis and Heat Treatment:

20.4 g of E. coli.
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118.1 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
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Sonication, output 8, duty 50/2 min, 7 times.
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Heat 15 min, 70 deg C.
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Centrifuge at 100,000 rcf, 50 min.
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134 ml of supernatant.
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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,2.0M AS 2.0M AS 0M AS 4 1.88M AS
2 Resource Q 6 ml 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 2M NaCl 4 Flow through
3 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 87min
4            
5            
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8            
9            

Comment:

Products were confirmed by MS.

2011.10.20

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