Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA0478
trehalose synthase

Lot Information:

Purification Lot: PC010478-41-01-03
sPlasmid ID: PC010478-41
Plasmid Vector: pET-11a
Label: Native
Person: Ishido E.
Purified Date: 02/15/2005
Total Protein Yield: 24.4 mg
Final Volume: 2.3 mL
Conc.: 10.6 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

01/18/2005Rosetta(DE3)6 L
01/26/2005Rosetta(DE3)6 L

Cell Lysis and Heat Treatment:

34.7g 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
ammonium sulfate precipitation20 mM Tris-HCl (pH 8.0), 50 mM NaCl, 1.5 M (NH4)2SO4  
Resource Q 6ml50 mM Tris-HCl (pH 9.0)5300 mM NaCl
Mono Q 8ml50 mM Tris-HCl (pH 9.0)4420 mM NaCl
Hydroxyapatite CHT5-I10 mM NaPi (pH 7.0), 150 mM NaCl450 mM NaPi
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.551 ml
HiPrep 26/10 Desalting 53ml20 mM Tris-HCl (pH 8.0)6