Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA0244
single-stranded DNA binding protein

Lot Information:

Purification Lot: PC010244-42-02-02
sPlasmid ID: PC010244-42
Plasmid Vector: pET-11a
Label: SeMet
Person: Ohmori M.
Purified Date: 09/17/2004
Total Protein Yield: 1.7 mg
Final Volume: 0.23 mL
Conc.: 7.3 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

08/26/2004B834(DE3)6 L

Cell Lysis and Heat Treatment:

19.1g 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)2SO450.57 M AS
Resource Q 6ml20 mM Tris-HCl (pH 8.0)3.50.30 M NaCl
Hydroxyapatite CHT2-I10 mM NaPi (pH 7.0), 150 mM NaCl20.03 M NaPi
HiTrap Heparin 5ml10 mM NaPi (pH 7.0)30.15 M NaCl
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.558 ml
HiPrep 26/10 Desalting 53ml20 mM Tris-HCl (pH 8.0)6