Whole Cell Project Database -Purification-

Show All

List View

 

Thermus thermophilus HB8  TTHA0654
ATP-binding protein, Mrp/Nbp35 family

Lot Information:

Purification Lot: PC010654-41-01-01
sPlasmid ID: PC010654-41
Plasmid Vector: pET-11a
Label: Native
Person: Kobayashi S.
Purified Date: 09/04/2001
Total Protein Yield:   mg
Final Volume:   mL
Conc.:   mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

08/09/2001BL21(DE3)4 L

Cell Lysis and Heat Treatment:

17.1g of E. coli cells
Ultra Sonication with  
Heat Treatment at  °C for   min
Ultra Cengrifuge at 200,000G for 1 hr

Purification Procedure:

ColumnBufferFlow Rate (mL/min)Eluted
Super Q Toyopearl 650M (PACKED)20 mM Tris-HCl (pH 8.0)7Flow through
0.01 M NaCl
Hydroxyapatite CHT10-I10 mM NaPi (pH 7.0)580 mM NaPi
Resource Q 6ml20 mM Tris-HCl (pH 8.0)30.2 - 0.5 M NaCl
Resource PHE 1ml50 mM NaPi (pH 7.0), 0.5 M (NH4)2SO410.13 M AS
38% Ethylene glycol
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.5