Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA0812
phosphoribosylamine-glycine ligase, PurD

Lot Information:

Purification Lot: PC010812-41-01-05
sPlasmid ID: PC010812-41
Plasmid Vector: pET-11a
Label: Native
Person: Nagira T.
Purified Date: 07/26/2007
Total Protein Yield: 98.5 mg
Final Volume: 6.66 mL
Conc.: 14.8 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

07/11/2007BL21(DE3)3 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), 1.5 M (NH4)2SO450.78M AS
Resource Q 6ml20 mM Tris-HCl (pH 8.0)569mM NaCl
Hydroxyapatite CHT10-I10 mM NaPi (pH 7.0), 150 mM NaCl4flow through
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.561.9ml