Mark J van Raaij
Inviato: sabato 1 aprile 2017 11.46
A: CCP4BB@JISCMAIL.AC.UK
Oggetto: Re: [ccp4bb] binding protein to nickel column
Could it be due to the fact that the protein is a histidine kinase - perhaps it
is interacting with or even modifying its own his-tag?
Mark J van Raaij
Dpto de
Could it be due to the fact that the protein is a histidine kinase - perhaps it
is interacting with or even modifying its own his-tag?
Mark J van Raaij
Dpto de Estructura de Macromoleculas
Centro Nacional de Biotecnologia - CSIC
calle Darwin 3
E-28049 Madrid, Spain
tel. (+34) 91 585 4616
http://w
Hi Shubhangi,
As Edward suggested, you can try with N-terminal
His tag. For this you can either clone in N-terminal His tagged based
vectors or by site directed insertion of 18 nucleotides coding for 6 His at
the N-terminus region.
But, before that, I suggest
K"
Subject: [ccp4bb] binding protein to nickel column
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
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 N
