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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species  
Locus tag APE0162
Products  
Plasmid ID 4079
Vector  
Purification Lot APE0162
Purified Date 2007.2.14-2.27
Protein conc. (mg/ml) 14.97
Total volume (ml) 0.87
Total protein (mg) 13.20

Culture Lot:

date host cell scale label weight of bacteira (g)
        40

Cell Lysis and Heat Treatment:

40 g of E. coli.
↓
200 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 150mM NaCl.
↓
Sonication output 8, duty 50/4min, 4 times.
↓
Heat 10 min, 80 deg C.
↓
Centrifuge at 100,000 rcf for 60 min.
↓
179 ml of supernatant.
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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,2M AS 2M AS 0M AS 0.9 1.45M AS
2 Resource Q 6 ml 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 1.2M NaCl 4 0.35M NaCl
3 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 77min
4            
5            
6            
7            
8            
9            

Comment:

Products were confirmed by Trp-MS.

2011.10.20

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