Whole Cell Project Database -Purification-

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

Lot Information:

Purification Lot: PC010926-41-02-03
sPlasmid ID: PC010926-41
Plasmid Vector: pET-11a
Label: SeMet
Person: Ebihara A.
Purified Date: 02/27/2002
Total Protein Yield: 9.3 mg
Final Volume: .8 mL
Conc.: 11.6 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

02/18/2002B834(DE3)6 L

Cell Lysis and Heat Treatment:

32.26g 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), 1.05 M (NH4)2SO45Flowthrough
Resource Q 6ml20 mM Tris-HCl (pH 8.0)5Flowthrough
Resource PHE 6ml50 mM NaPi (pH 7.0), 1.5 M (NH4)2SO450.55 M AS
Resource Q 6ml20 mM Tris-HCl (pH 8.0)50.05 M NaCl
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.573 ml