Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA0656
ribosomal protein L11 methyltransferase

Lot Information:

Purification Lot: PC010656-41-01-03
sPlasmid ID: PC010656-41
Plasmid Vector: pET-11a
Label: Native
Person: Maoka N.
Purified Date: 09/03/2002
Total Protein Yield: 78.5 mg
Final Volume: 3.4 mL
Conc.: 23.1 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

08/26/2002BL21(DE3)12 L

Cell Lysis and Heat Treatment:

26.8g 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
ammonium sulfate precipitation20 mM Tris-HCl (pH 8.0), 50 mM NaCl, 1.5 M (NH4)2SO4  
Resource Q 6ml20 mM Tris-HCl (pH 8.0)50.16 M NaCl
Hydroxyapatite CHT10-I10 mM NaPi (pH 7.0)4Flowthrough
Mono Q 8ml20 mM Tris-HCl (pH 8.0)50.19 M NaCl
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.569 ml