Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA1742
orotate phosphoribosyltransferase, pyrE

Lot Information:

Purification Lot: PC011742-41-01-01
sPlasmid ID: PC011742-41
Plasmid Vector: pET-11a
Label: Native
Person: Hoseki J.
Purified Date: 05/20/2002
Total Protein Yield: 7.8 mg
Final Volume: 0.9 mL
Conc.: 8.7 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

05/13/2002BL21(DE3)6 L

Cell Lysis and Heat Treatment:

14g 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
Resource PHE 6ml50 mM NaPi (pH 7.0), 1.2 M (NH4)2SO450.52 M ammonium sulfate
Resource Q 6ml20 mM Tris-HCl (pH 8.0)50.15 M NaCl
Hydroxyapatite CHT2-I10 mM NaPi (pH 7.0)2flowthrough
HiLoad 16/60 Superdex 200 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.591 ml