Hello, I have recently began using the Anti-NuMA primary antibody to distinguish the human mesechymal stem cells that I am delivering to the rat heart. Anti-NuMA recognizes human nuclear mitotic apparatus protein. I have tried two secondary antibodies: Alexa Fluor 488 and FITC (Jackson immunoresearch code 715-095-150) and have positively stained human mesenchymal stem cells put on glass coverslips with both secondary antibodies. However, when I look at the sections of my rat heart that I have delivered human mesenchymal stem cells to, I get a lot of autofluorescence and no positive signal (counterstained with Hoechst and the hMSCs are Quantum Dot loaded). The sections were fixed in 4% paraformaldehyde right after the animal was euthanized and heart removed. The heart was then frozen in OCT and sectioned. I have tried changing the concentration of both the secondary's and primary's already (within the manufacturer's specs). My question is: what secondary antibodies do you recommend? Does anyone have experience with this primary?
Thanks, Sharon WPI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
