Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA1199
ornithine carbamoyltransferase

Lot Information:

Purification Lot: PC011199-41-01-02
sPlasmid ID: PC011199-41
Plasmid Vector: pET-11a
Label: Native
Person: Nishida M.
Purified Date: 08/11/2006
Total Protein Yield: 3 mg
Final Volume: 0.3 mL
Conc.: 10.1 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

07/09/2006Rosetta(DE3)6 L

Cell Lysis and Heat Treatment:

21.7g of E. coli cells
Ultra Sonication with 20mM Tris-HCl + 50mM NaCl (pH8.0)
Heat Treatment at none°C for   min
Ultra Cengrifuge at 200,000G for 1 hr

Purification Procedure:

ColumnBufferFlow Rate (mL/min)Eluted
Resource ISO 6ml50 mM NaPi (pH 7.0), 1.5 M (NH4)2SO45flowthrough
Resource ISO 6ml50 mM NaPi (pH 7.0), 1.5 M (NH4)2SO450.78 M AS
Resource Q 6ml20 mM Tris-HCl (pH 8.0)560 mM NaCl
Hydroxyapatite CHT5-I10 mM NaPi (pH 7.0), 150 mM NaCl370 mM NaPi
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.552 mL