Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA0255
ferric uptake regulation protein

Lot Information:

Purification Lot: PC010255-41-01-04
sPlasmid ID: PC010255-41
Plasmid Vector: pET-11a
Label: Native
Person: Okada M.
Purified Date: 09/04/2007
Total Protein Yield: .9 mg
Final Volume: 0.153 mL
Conc.: 5.6 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

08/21/2007BL21(DE3)18 L

Cell Lysis and Heat Treatment:

46.6g of E. coli cells
Ultra Sonication with 20mM Tris-HCl + 50mM NaCl (pH8.0) + 5mM 2-ME
Heat Treatment at 70°C for 10 min
Ultra Cengrifuge at 200,000G for 1 hr

Purification Procedure:

ColumnBufferFlow Rate (mL/min)Eluted
HiPrep 26/10 Desalting 53ml20 mM Tris-HCl (pH 8.0)10 
Super Q Toyopearl 650M 80 ml20 mM Tris-HCl (pH 8.0)80.13M NaCl
Resource Q 6ml20 mM Tris-HCl (pH 8.0)40.02M NaCl
Resource ISO 1ml50 mM NaPi (pH 7.0), 1.5 M (NH4)2SO421.5M AS
Resource PHE 1ml50 mM NaPi (pH 7.0), 1.5 M (NH4)2SO421.26 AS
Resource PHE 1ml50 mM NaPi (pH 7.0), 1.5 M (NH4)2SO421.13M AS
HiLoad 16/60 Superdex 200 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.582.6 ml