Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA1515
phosphoribosylaminoimidazole-succinocarboxamide synthase

Lot Information:

Purification Lot: PC011515-41-02-05
sPlasmid ID: PC011515-41
Plasmid Vector: pET-11a
Label: SeMet
Person: Inoue Y.
Purified Date: 10/11/2007
Total Protein Yield:   mg
Final Volume:   mL
Conc.:   mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

08/02/2007B834(DE3)9 L

Cell Lysis and Heat Treatment:

27.7g 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
Ammonium sulfate precipitation20 mM Tris-HCl (pH 8.0), 50 mM NaCl, 1.5 M (NH4)2SO4 ppt
Resource Q 6ml20 mM Tris-HCl (pH 8.0)50.04M NaCl
Hydroxyapatite CHT10-I10 mM NaPi (pH 7.0)460mM NaPi
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.51:48.9ml
2:58.8ml