Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA0926
dephospho-CoA kinase (dephosphocoenzyme A kinase)

Lot Information:

Purification Lot: PC010926-41-02-04
sPlasmid ID: PC010926-41
Plasmid Vector: pET-11a
Label: SeMet
Person: Ebihara A.
Purified Date: 07/24/2002
Total Protein Yield:   mg
Final Volume:   mL
Conc.:   mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

07/10/2002B834(DE3)6 L

Cell Lysis and Heat Treatment:

31.2g 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)2SO45flowthrough
Resource PHE 6ml50 mM NaPi (pH 7.0), 1.5 M (NH4)2SO450.7 M AS
Resource Q 6ml20 mM Tris-HCl (pH 8.0)50.02 and 0.07 M NaCl
Mono Q 1ml20 mM Tris-HCl (pH 8.0)10.03 (A) and 0.06 (B) M NaCl
Mono Q 1ml20 mM Tris-HCl (pH 8.0)10.03 (A) and 0.04 (B) M NaCl
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.5