Size Exclusion Column - Proteins not coming out ! - BMP-2 (30 kDa) is inhjected in a colum (size exclusion at 100kDa) and (Nov/26/2007 )
Hello all,
I can't seem to understand why my protein ( BMP-2 in 1mM HCl) does not come out after injecting it on a size exclusion column. I have eluded with 1mM HCl (45ml, void volume is 10ml) but I don't have any sign of the protein... It must stick somewhere in the column... How can I prevent this from happening ?
-Coockie-
What type of column was it?
Protein concentration used?
Do you have an issue with including 150 mM NaCl to the buffer?
-genehunter-1-
QUOTE (Coockie @ Nov 26 2007, 01:16 PM)
Hello all,
I can't seem to understand why my protein ( BMP-2 in 1mM HCl) does not come out after injecting it on a size exclusion column. I have eluded with 1mM HCl (45ml, void volume is 10ml) but I don't have any sign of the protein... It must stick somewhere in the column... How can I prevent this from happening ?
I can't seem to understand why my protein ( BMP-2 in 1mM HCl) does not come out after injecting it on a size exclusion column. I have eluded with 1mM HCl (45ml, void volume is 10ml) but I don't have any sign of the protein... It must stick somewhere in the column... How can I prevent this from happening ?
may be unspecifically bound to the matrix; you may block with BSA (if you accept impurities) or use some triton x-100
-The Bearer-