Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHB149
CRISPR-associated protein Csm3

Lot Information:

Purification Lot: PC014149-42-01-01
sPlasmid ID: PC014149-42
Plasmid Vector: pET-HisTEV
Label: Native
Person: Takahara M.
Purified Date: 10/12/2007
Total Protein Yield: .6 mg
Final Volume: 0.355 mL
Conc.: 1.7 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

09/13/2007BL21(DE3)18 L

Cell Lysis and Heat Treatment:

51.7g of E. coli cells
Ultra Sonication with 20mM Tris-HCl + 500mM NaCl (pH8.0) + 5mM 2-ME
Heat Treatment at 70°C for 10 min
Ultra Cengrifuge at 200,000G for 1 hr

Purification Procedure:

ColumnBufferFlow Rate (mL/min)Eluted
HisTrap HP 5ml20 mM Tris-HCl (pH 8.0), 0.5 M NaCl, 20 mM Imidazole50.5M Imidazole
HiLoad 26/60 Superdex 200 pg20 mM Tris-HCl (pH 8.0), 200 mM NaCl, 20 mM Imidazole1.588ml
Protease Treatment  OK
HisTrap HP 5ml20 mM Tris-HCl (pH 8.0), 0.5 M NaCl, 20 mM Imidazole5Flowthrough
HiPrep 26/10 Desalting 53ml20 mM Tris-HCl (pH 8.0), 200 mM NaCl7