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Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST0212
Products  
Plasmid ID 0149
Vector  
Purification Lot ST0212_1
Purified Date 2004.9.13
Protein conc. (mg/ml) -
Total volume (ml) -
Total protein (mg) -

Culture Lot:

date host cell scale label weight of bacteira (g)
        13

Cell Lysis and Heat Treatment:

 
 
 
 
 
 
 
 
 

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,1.08M AS 1.08M AS 0M AS 5 600mM AS
2 Resource S 6mL 50mM MES(pH6.0),5mM B-ME,120mM NaCl 120mM NaCl 700mM NaCl 5 flowthrough and 200mM NaCl
3 HiTrap Heparin HP 50mM MES(pH6.0),5mM B-ME,120mM NaCl 120mM NaCl 800mM NaCl 3 460mM NaCl
4 Bio-Scale CHT 10I 10mM KPi(pH7.0),5mM B-ME 10mM KPi 500mM KPi 3 240mM KPi
5 Resource Q 6mL 20mM Tris-HCl(pH9.0),5mM B-ME 0M NaCl 500mM NaCl 5 150mM NaCl
6 BioAssist Q 10X10 20mM Tris-HCl(pH9.0),5mM B-ME 0M NaCl 500mM NaCl 3 110mM NaCl
7 HiTrap Blue HP 20mM Tris-HCl(pH9.0),5mM B-ME 0M NaCl 800mM NaCl 3 800mM NaCl
8 HiLoad 16/60 Superdex75 50mM Tris-HCl(pH9.0),5mM B-ME,150mM NaCl     1  
9            

Comment:

It was hard to solve this solution at less than 110 mM NaCl or less than pH9.0. Solution was yellow.

2011.10.20

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