Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA0244
single-stranded DNA binding protein

Lot Information:

Purification Lot: PC010244-42-01-04
sPlasmid ID: PC010244-42
Plasmid Vector: pET-11a
Label: Native
Person: Nagira T.
Purified Date: 06/19/2007
Total Protein Yield:   mg
Final Volume:   mL
Conc.:   mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

05/31/2007BL21(DE3)9 L

Cell Lysis and Heat Treatment:

30.6g 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)2SO450.64M AS
Resource Q 6ml20 mM Tris-HCl (pH 8.0)51:308mM NaCl
2:464mM NaCl
HiTrap Heparin 5ml10 mM NaPi (pH 7.0)41:149mM NaCl
2:flow through
Hydroxyapatite CHT5-I10 mM NaPi (pH 7.0), 150 mM NaCl31:37mM NaPi
2:46mM NaPi
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.51:54.0ml
2:46.8ml
HiPrep 26/10 Desalting 53ml20 mM Tris-HCl (pH 9.0), 150 mM NaCl6