Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHB054
precorrin-2 methylase

Lot Information:

Purification Lot: PC014054-41-01-02
sPlasmid ID: PC014054-41
Plasmid Vector: pET-11a
Label: Native
Person: Agari Y.
Purified Date: 03/25/2002
Total Protein Yield: 19.3 mg
Final Volume: 1.27 mL
Conc.: 15.2 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

03/14/2002BL21(DE3)6 L

Cell Lysis and Heat Treatment:

17.3g 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)2SO45660mM (NH4)2SO4
Hydroxyapatite CHT10-I10 mM NaPi (pH 7.0)564mM NaPi
Resource Q 6ml20 mM Tris-HCl (pH 8.0)5130mM NaCl
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.564.5 ml