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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species Aeropyrum pernix
Locus tag APE0317
Products phosphomannomutase
Plasmid ID R4081
Vector  
Purification Lot APE0317_1
Purified Date 2005.11.21-12.2
Protein conc. (mg/ml) 7.18
Total volume (ml) 1.00
Total protein (mg) 7.18

Culture Lot:

date host cell scale label weight of bacteira (g)
        116

Cell Lysis and Heat Treatment:

116 g of E. coli.
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ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 150mM NaCl.
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Sonication, output 8, duty 50/2min, 5 times.
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Heat 15 min, 85 deg C.
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Centrifuge at 100,000 rcf for 60 min.
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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,1.05M AS 1.05M AS 0M AS 4 0.8M AS
2 Resource Q 6 ml 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 0.5M NaCl 3 0.25M NaCl
3 HiLoad 16/60 Superdex200 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 65ml/120ml
4            
5            
6            
7            
8            
9            

Comment:

Products were confirmed by MS.

2011.10.20

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