Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA1394
homoserine kinase

Lot Information:

Purification Lot: PC011394-41-01-01
sPlasmid ID: PC011394-41
Plasmid Vector: pET-11a
Label: Native
Person: Maoka N.
Purified Date: 06/01/2001
Total Protein Yield: 16.8 mg
Final Volume: 2.2 mL
Conc.: 7.6 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

05/10/2001BL21(DE3)2 L

Cell Lysis and Heat Treatment:

9.9g 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 ISO 6ml50 mM NaPi (pH 7.0), 1.2 M (NH4)2SO450.29 M AS
Resource Q 6ml20 mM Tris-HCl (pH 8.0)50.11 M NaCl
Hydroxyapatite CHT2-I10 mM NaPi (pH 7.0)3Flowthrough
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.559 ml