Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHB054
precorrin-2 methylase

Lot Information:

Purification Lot: PC014054-41-02-03
sPlasmid ID: PC014054-41
Plasmid Vector: pET-11a
Label: SeMet
Person: Ebihara A.
Purified Date: 09/13/2002
Total Protein Yield: .6 mg
Final Volume: 1.4 mL
Conc.: .4 mg/mL


Culture Lot:


host cell



07/03/2002B834(DE3)6 L

Cell Lysis and Heat Treatment:

18.4g of E. coli cells
Ultra Sonication with 20mM MES + 50mM NaCl (pH6.0) + 10% Glycerol
Heat Treatment at 70°C for 10 min
Ultra Cengrifuge at 200,000G for 1 hr

Purification Procedure:

ColumnBufferFlow Rate (mL/min)Eluted
Resource ISO 6ml50 mM NaPi (pH 7.0), 1.35 M (NH4)2SO450.6 M AS
Hydroxyapatite CHT2-I10 mM NaPi (pH 7.0)240 mM NaPi
Resource Q 6ml20 mM Tris-HCl (pH 8.0)50.1 M NaCl
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.5