Whole Cell Project Database -Purification-

Show All

List View

 

Thermus thermophilus HB8  TTHA0255
ferric uptake regulation protein

Lot Information:

Purification Lot: PC010255-41-01-05
sPlasmid ID: PC010255-41
Plasmid Vector: pET-11a
Label: Native
Person: Inoue Y.
Purified Date: 07/11/2008
Total Protein Yield: 12 mg
Final Volume: 0.7 mL
Conc.: 17.1 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

08/21/2007BL21(DE3)18 L

Cell Lysis and Heat Treatment:

31.0g 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
Super Q Toyopearl 650M (PACKED)20 mM Tris-HCl (pH 8.0)5flowthrough
HiTrap Heparin 5ml50 mM Tris-HCl (pH 9.0), 100 mM NaCl30.62 M NaCl
Resource Q 6ml50 mM Tris-HCl (pH 9.0), 100 mM NaCl5flowthrough ~ 0.15 M NaCl
Resource PHE 6ml50 mM NaPi (pH 7.0), 2.5 M (NH4)2SO451.39 M AS
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 0.15 M NaCl0.557.6 ml