Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA1969
chromosome partitioning protein, ParB family

Lot Information:

Purification Lot: PC011969-42-01-05
sPlasmid ID: PC011969-42
Plasmid Vector: pET-11a
Label: Native
Person: Nishida M.
Purified Date: 09/13/2005
Total Protein Yield: 15.4 mg
Final Volume: 1.02 mL
Conc.: 15.1 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

06/30/2005Rosetta(DE3)4.5 L

Cell Lysis and Heat Treatment:

15.1g of E. coli cells
Ultra Sonication with 20mM Tris-HCl + 500mM 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
HiPrep 26/10 Desalting 53ml20 mM Tris-HCl (pH 8.0)10 
Super Q Toyopearl 650M 80 ml20 mM Tris-HCl (pH 8.0)8130 mM NaCl
Resource Q 6ml20 mM Tris-HCl (pH 8.0)4flowthrough
Hydroxyapatite CHT10-I10 mM KPi (pH 7.0)3180 mM KPi
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 200 mM NaCl0.555 mL