Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA1969
chromosome partitioning protein, ParB family

Lot Information:

Purification Lot: PC011969-42-02-06
sPlasmid ID: PC011969-42
Plasmid Vector: pET-11a
Label: SeMet
Person: Inoue Y.
Purified Date: 12/01/2005
Total Protein Yield: .3 mg
Final Volume: 0.11 mL
Conc.: 2.8 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

10/07/2005Rosetta834(DE3)12 L

Cell Lysis and Heat Treatment:

27.1g 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)6 
Super Q Toyopearl 650M 30 ml20 mM Tris-HCl (pH 8.0)40.19M NaCl
Resource Q 6ml20 mM Tris-HCl (pH 8.0)40.07M NaCl
Hydroxyapatite CHT5-I10 mM KPi (pH 7.0)2220mM KPi
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 200 mM NaCl0.552.7ml