Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA0478
trehalose synthase

Lot Information:

Purification Lot: PC010478-41-01-04
sPlasmid ID: PC010478-41
Plasmid Vector: pET-11a
Label: Native
Person: Nishida M.
Purified Date: 09/15/2005
Total Protein Yield: 3.2 mg
Final Volume: 0.26 mL
Conc.: 12.2 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

06/30/2005Rosetta(DE3)4.5 L

Cell Lysis and Heat Treatment:

15.2g of E. coli cells
Ultra Sonication with 20mM Tris-HCl + 500mM 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
HiPrep 26/10 Desalting 53ml20 mM Tris-HCl (pH 8.0)10 
Super Q Toyopearl 650M 80 ml20 mM Tris-HCl (pH 8.0)8190 mM NaCl
Resource Q 6ml20 mM Tris-HCl (pH 8.0)4260 mM NaCl
Hydroxyapatite CHT10-I10 mM KPi (pH 7.0)3flowthrough
HiLoad 16/60 Superdex 200 pg20 mM Tris-HCl (pH 8.0), 200 mM NaCl0.552 mL
HiPrep 26/10 Desalting 53ml20 mM Tris-HCl (pH 8.0)10