Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA1427
UvrD protein

Lot Information:

Purification Lot: PC011427-41-02-02
sPlasmid ID: PC011427-41
Plasmid Vector: pET-11a
Label: SeMet
Person: Nishida M.
Purified Date: 09/12/2007
Total Protein Yield:   mg
Final Volume:   mL
Conc.:   mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

09/07/2007Rosetta834(DE3)18 L

Cell Lysis and Heat Treatment:

25.0g 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)2SO450.44 M AS
Resource Q 6ml20 mM Tris-HCl (pH 8.0), 150 mM NaCl50.22 M NaCl
HiTrap Heparin 5ml20 mM Tris-HCl (pH 8.0), 150 mM NaCl40.27 M NaCl
HiLoad 16/60 Superdex 200 pg20 mM Tris-HCl (pH 8.0), 0.5 M NaCl0.5A: 59 mL
B: 71 mL