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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species Sulfolobus tokodaii
Locus tag ST1895
Products hypothetical glycine cleavage system H protein
Plasmid ID R0471
Vector  
Purification Lot ST1895
Purified Date 2006.2.7-2.15
Protein conc. (mg/ml) 33.55
Total volume (ml) 3.75
Total protein (mg) 125.8

Culture Lot:

date host cell scale label weight of bacteira (g)
        13.8

Cell Lysis and Heat Treatment:

13.8 g of E. coli.
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90ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
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Sonication, output 8, duty 50/3 min, 5 times.
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Heat 20 min, 75 deg C.
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Centrifuge at 100,000 rcf, 60 min.
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66 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,1.5M AS 1.5M AS 0M AS 6 Flow through
2 Resource Phe 6ml 20mM Tris-HCl(pH8.0),5mM B-ME,2.0M AS 2.0M AS 0M AS 6 Flow through
3 Resource S 6 ml 50mM MES(pH6.0),5mM B-ME 0M NaCl 2M NaCl 4 from 0.10M NaCl to 0.150M NaCl
4 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 87min
5            
6            
7            
8            
9            

Comment:

Products were confirmed by Trp-Mass.

2011.10.20

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