Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA1969
chromosome partitioning protein, ParB family

Lot Information:

Purification Lot: PC011969-42-02-02
sPlasmid ID: PC011969-42
Plasmid Vector: pET-11a
Label: SeMet
Person: Manzoku M.
Purified Date: 10/15/2004
Total Protein Yield: 4.9 mg
Final Volume: 0.20 mL
Conc.: 24.6 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

09/03/2004Rosetta834(DE3)3 L
09/03/2004Rosetta834(DE3)3 L

Cell Lysis and Heat Treatment:

15.9g 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)2SO450.67M AS
Hydroxyapatite CHT5-I10 mM NaPi (pH 7.0), 0.5 M NaCl3flow through
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 0.5 M NaCl0.550.5 mL