IFA of cell surface markers - IFA of osteoclasts not working, help! (Nov/20/2008 )

Hi all,

I'm having quite a few problems with my IFA (immunofluorescence assay) of osteoclasts. I've tried fixing with PFA or MeOH and I still have faint staining when it should light up like a christmas tree. The markers I'm staining for are on the cell surface and I'm wondering if I still have to permeabilize the cells or if I should just fix them, and if so which fixative works best for this? Thanks a bunch!!

Dana

Hi all,

I'm having quite a few problems with my IFA (immunofluorescence assay) of osteoclasts. I've tried fixing with PFA or MeOH and I still have faint staining when it should light up like a christmas tree. The markers I'm staining for are on the cell surface and I'm wondering if I still have to permeabilize the cells or if I should just fix them, and if so which fixative works best for this? Thanks a bunch!!

Dana

fantastic! thank you sooo much!!

dana

QUOTE (danalcat @ Nov 20 2008, 08:30 AM)

Hi all,

I'm having quite a few problems with my IFA (immunofluorescence assay) of osteoclasts. I've tried fixing with PFA or MeOH and I still have faint staining when it should light up like a christmas tree. The markers I'm staining for are on the cell surface and I'm wondering if I still have to permeabilize the cells or if I should just fix them, and if so which fixative works best for this? Thanks a bunch!!

Dana

May be this article will help.