Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA0656
ribosomal protein L11 methyltransferase

Lot Information:

Purification Lot: PC010656-41-02-02
sPlasmid ID: PC010656-41
Plasmid Vector: pET-11a
Label: SeMet
Person: Maoka N.
Purified Date: 03/06/2002
Total Protein Yield: 15.1 mg
Final Volume: 1.15 mL
Conc.: 13.1 mg/mL


Culture Lot:


host cell



02/28/2002B834(DE3)4 L

Cell Lysis and Heat Treatment:

13g 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 precipitation50 mM NaPi (pH 7.0), 1.5 M (NH4)2SO4  
Resource Q 6ml20 mM Tris-HCl (pH 8.0)50.17 M NaCl
Hydroxyapatite CHT2-I10 mM NaPi (pH 7.0)2Flowthrough
BioAssist Q 0.83ml20 mM Tris-HCl (pH 8.0)0.50.28 M NaCl
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.5