Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA0606
aspartate 1-decarboxylase

Lot Information:

Purification Lot: PC010606-41-01-01
sPlasmid ID: PC010606-41
Plasmid Vector: pET-11a
Label: Native
Person: Maoka N.
Purified Date: 09/11/2001
Total Protein Yield: 108.2 mg
Final Volume: 4.7 mL
Conc.: 23 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

08/29/2001BL21(DE3)4 L

Cell Lysis and Heat Treatment:

14.72g 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)2SO450.68 M AS
Resource Q 6ml20 mM Tris-HCl (pH 8.0)50.07 M NaCl
Hydroxyapatite CHT10-I10 mM NaPi (pH 7.0)4Flowthrough
Resource S 6ml20 mM MES (pH 6.0) 5Flowthrough
Resource Q 6ml20 mM Tris-HCl (pH 8.0)50.07 M NaCl
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.564 ml