Immunostaining of peroxysome - (Jun/06/2005 )
does anyone ever did an IHC / ICC / IF of peroxysome either in live cells or on fixed cells?
My specific questions are:
1) I got a staining in transfected, live cells that makes me thinking of peroxysome-like, but I never tried a double staining
=> what good antibody to use ?... (rat / mouse / human fibro cell lines)
2) I think I got a kind of swelling of these structures when fixing cells. Does anybody ever remarked this?
Thanks for replies!!
Hi ... how did you fix your preps first off which may explain the swelling?
Are you looking for a specific peroxisome marker? I don't specifically have experience with such a thing but did a search and came up with PMP 70 and catalase as being markers. I see several publications using these.
If you are looking for an antibody that stains rat, mouse and human fibro cells lines I would think you probably need a polyclonal but maybe not. Otherwise you could purchase a few different monos for your various cell lines.
Also do you have a pic of what your first staining looked like. Was it done by IHC or IF, antibody based protocol? This may help if you want to design a double staining procedure.
Hope it helps some and let me know if you have more info as perhaps we could help design a staining protocol!