Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA1427
UvrD protein

Lot Information:

Purification Lot: PC011427-41-01-01
sPlasmid ID: PC011427-41
Plasmid Vector: pET-11a
Label: Native
Person: Inoue Y.
Purified Date: 06/13/2007
Total Protein Yield:   mg
Final Volume:   mL
Conc.:   mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

05/30/2007Rosetta(DE3)9 L

Cell Lysis and Heat Treatment:

32.3g of E. coli cells
Ultra Sonication with 20mM Tris-HCl + 50mM NaCl (pH8.0)
Heat Treatment at 55°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)2SO450.48M AS
Resource Q 6ml20 mM Tris-HCl (pH 8.0), 150 mM NaCl50.22 NaCl
HiTrap Heparin 5ml10 mM NaPi (pH 7.0), 150 mM NaCl30.35M NaCl
HiLoad 16/60 Superdex 200 pg20 mM Tris-HCl (pH 8.0), 0.5 M NaCl0.51:72.0ml
2:82.8ml