Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA1027
DNA integration/recombination/invertion protein

Lot Information:

Purification Lot: PC011027-41-02-01
sPlasmid ID: PC011027-41
Plasmid Vector: pET-11a
Label: SeMet
Person: Shinkai F.
Purified Date: 03/11/2005
Total Protein Yield: 4.4 mg
Final Volume: 0.7 mL
Conc.: 6.3 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

05/14/2004Rosetta834(DE3)6 L
01/14/2005BL21(DE3)6 L

Cell Lysis and Heat Treatment:

44.8g 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 PHE 6ml50 mM NaPi (pH 7.0), 1.5 M (NH4)2SO450.58 M AS
HiTrap Heparin 5ml10 mM NaPi (pH 7.0), 0.5 M NaCl41.0 M NaCl
HiLoad 16/60 Superdex 200 pg20 mM Tris-HCl (pH 8.0), 0.5 M NaCl0.577 ml