Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA1642
S-adenosylmethionine synthetase

Lot Information:

Purification Lot: PC011642-42-01-01
sPlasmid ID: PC011642-42
Plasmid Vector: pET-11a
Label: Native
Person: Shinkai F.
Purified Date: 06/10/2005
Total Protein Yield: 150.8 mg
Final Volume: 8 mL
Conc.: 18.9 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

04/12/2005Rosetta(DE3)6 L

Cell Lysis and Heat Treatment:

23.3g of E. coli cells
Ultra Sonication with 20mM Tris-HCl + 50mM NaCl (pH8.0)
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), 1.5 M (NH4)2SO450.64 M AS
Resource Q 6ml20 mM Tris-HCl (pH 8.0)50.15 M NaCl
Hydroxyapatite CHT10-I10 mM NaPi (pH 7.0), 150 mM NaCl440 mM NaPi
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.550 ml
HiPrep 26/10 Desalting 53ml20 mM Tris-HCl (pH 8.0)6