Hello
I have a 10kda histidine kinase domain protein with a pI of 9.5.
It has a C-term his tag and despite using different buffers the protein
doesnt bind to the nickel cloumn. it comes out in the flow trhough.
Buffers used- 50mM tris Ph=8, 300mM NaCl
50mM tris Ph=7, 300mM NaCl
50mM hepes Ph=7.5, 300mM NaCl
50mM tris Ph=8, 300mM NaCl, 10% glycerol
Can someone suggest to get ensure binding of the protein to the nickel-nta
column
Shubhangi
PhD student
under Dr. Jhimli Dasgupta
St. Xavier's College
Kolkata
India
