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Plasmid & Protein Purification Database -Purification-

Species  
Locus tag STS042
Products  
Plasmid ID 0155
Vector  
Purification Lot STS042_3a
Purified Date 2005.5.13-5.24
Protein conc. (mg/ml) 15.5
Total volume (ml) 0.93
Total protein (mg) 14.4

Culture Lot:

date host cell scale label weight of bacteira (g)
        19.6

Cell Lysis and Heat Treatment:

19.6 g of E. coli.
↓
100 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 150mM NaCl.
↓
Sonication, output 8/duty 5, 2 min, 5 times.
↓
Heat 15min, 75 deg C.
↓
Centrifuge at 100,000 rcf for 50min.
↓
90 ml of supernatant.
↓
 
↓
 
 

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,2.0M AS 2.0M AS 0M AS 6 from 1.93M AS to 1.58M AS
2 BioAssist Q 10x10 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 0.25M NaCl 2 from 0.02M NaCl to 0.04M NaCl
3 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 60ml/120ml
4            
5            
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8            
9            

Comment:

Products were devided in the gel filtration step into two peaks, the main peak (a, possibly multimer) and the sub peak (b, possibly monomer). Refer the previous purification lot.

2011.10.20

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