Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA0512
hypothetical protein

Lot Information:

Purification Lot: PC010512-41-02-03
sPlasmid ID: PC010512-41
Plasmid Vector: pET-11a
Label: SeMet
Person: Nagira T.
Purified Date: 10/19/2005
Total Protein Yield: .4 mg
Final Volume: 0.06 mL
Conc.: 6.3 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

09/27/2005B834(DE3)6 L

Cell Lysis and Heat Treatment:

18.8g 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), 1.5 M (NH4)2SO45277mM AS
Resource Q 6ml20 mM Tris-HCl (pH 8.0)5170mM NaCl
Hydroxyapatite CHT2-I10 mM NaPi (pH 7.0), 150 mM NaCl296mM NaPi
HiLoad 16/60 Superdex 200 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.560ml
HiPrep 26/10 Desalting 53ml20 mM Tris-HCl (pH 8.0)6