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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species  
Locus tag APE0519
Products  
Plasmid ID 4024
Vector  
Purification Lot APE0519_2
Purified Date 2005.11.10-11.29
Protein conc. (mg/ml)  
Total volume (ml)  
Total protein (mg)  

Culture Lot:

date host cell scale label weight of bacteira (g)
        42

Cell Lysis and Heat Treatment:

42 g of E. coli.
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200 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 20 min, 80 deg C.
↓
Centrifuge at 100,000 rcf for 45 min.
↓
184 (200) 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 0.9M AS 6 from 1.25M AS to 1.1M AS
2 Resource S 6 ml 50mM MES(pH6.0),5mM B-ME 0M NaCl 0.12M NaCl 4 0.05M NaCl
3 Bio-Scale CHT2I 10mM KPi(pH7.0),5mM B-ME 10mM Kpi 200mM Kpi 3 0.1M Kpi
4 HiLoad 16/60 Superdex 75          
5            
6            
7            
8            
9            

Comment:

 

2011.10.20

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