Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA0812
phosphoribosylamine-glycine ligase, PurD

Lot Information:

Purification Lot: PC010812-41-02-03
sPlasmid ID: PC010812-41
Plasmid Vector: pET-11a
Label: SeMet
Person: Maoka N.
Purified Date: 07/30/2002
Total Protein Yield: 39.6 mg
Final Volume: 1.6 mL
Conc.: 24.7 mg/mL


Culture Lot:


host cell



07/17/2002B834(DE3)6 L

Cell Lysis and Heat Treatment:

37.7g 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), 1.35 M (NH4)2SO450.70 M AS
Resource Q 6ml20 mM Tris-HCl (pH 8.0)50.12 M NaCl
Hydroxyapatite CHT2-I10 mM NaPi (pH 7.0), 150 mM NaCl2Flowthrough
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.564 ml