apolipoprotein E identification via Westerns - (Mar/10/2005 )
Hello friends,
I am currently working on identifying expression of apo E on pulmonary artery smooth muscle cells via Invitrogen NuPAGE WB. One thing I am finding is that my monoclonal ab is identifying a band around 65-70 KDa in size in some bands which I think may be a complex of apo E monomers.
Has anyone seen something like that before?
What would you advise as the best strategy to further reduce the sample and isolate monomers that would go with my positive controls?
Thanks for any feedback you can offer. All the best, V
-ubiquitroni-
QUOTE (ubiquitroni @ Mar 10 2005, 09:22 PM)
Hello friends,
I am currently working on identifying expression of apo E on pulmonary artery smooth muscle cells via Invitrogen NuPAGE WB. One thing I am finding is that my monoclonal ab is identifying a band around 65-70 KDa in size in some bands which I think may be a complex of apo E monomers.
Has anyone seen something like that before?
What would you advise as the best strategy to further reduce the sample and isolate monomers that would go with my positive controls?
Thanks for any feedback you can offer. All the best, V
I am currently working on identifying expression of apo E on pulmonary artery smooth muscle cells via Invitrogen NuPAGE WB. One thing I am finding is that my monoclonal ab is identifying a band around 65-70 KDa in size in some bands which I think may be a complex of apo E monomers.
Has anyone seen something like that before?
What would you advise as the best strategy to further reduce the sample and isolate monomers that would go with my positive controls?
Thanks for any feedback you can offer. All the best, V
I have had this same problem with Apo-AI, I get what appears to be a dimer from 65-70KDa. I tried different concentration of BME and DTT but no luck. Next week I'm going to run again with a positive and negative control and a new antibody (not like the antibody would really help). I can't for the life of me get rid of the dimer. I may try DTT in the lysis buffer. I can't get it to go away either!
-ScienceBoy81-