Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA1515
phosphoribosylaminoimidazole-succinocarboxamide synthase

Lot Information:

Purification Lot: PC011515-41-01-02
sPlasmid ID: PC011515-41
Plasmid Vector: pET-11a
Label: Native
Person: Maoka N.
Purified Date: 11/07/2001
Total Protein Yield: 112.1 mg
Final Volume: 8.4 mL
Conc.: 13.3 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

10/31/2001BL21(DE3)6 L

Cell Lysis and Heat Treatment:

17.75g of E. coli cells
Ultra Sonication with  
Heat Treatment at  °C for   min
Ultra Cengrifuge at 200,000G for 1 hr

Purification Procedure:

ColumnBufferFlow Rate (mL/min)Eluted
Ammonium sulfate precipitation50 mM NaPi (pH 7.0), 1.5 M (NH4)2SO4  
Resource Q 6ml20 mM Tris-HCl (pH 8.0)5Flowthrough ~ 0.04 M NaCl
Hydroxyapatite CHT10-I10 mM NaPi (pH 7.0)440 ~ 80 mM NaPi
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.5