CD31 Staining of PFA fixed Frozen Tissue - (Feb/27/2008 )
Hello all,
Any recommendations for immunhistochemistry protocols for CD31 staining? I am currently trying it on tissues which have been fixed in 4% PFA and sectioned on a cryostat, using the MEC13.3 antibody (BD Bioscience) which I understand is the best. I'm not getting very good staining though, and am wondering if the PFA fixation is interfering with antibody binding (they do not recommend this antibody for formalin fixed paraffin embedded tissues). Any suggestions?
Mike
-dankasfuk-
QUOTE (dankasfuk @ Feb 27 2008, 04:09 PM)
Hello all,
Any recommendations for immunhistochemistry protocols for CD31 staining? I am currently trying it on tissues which have been fixed in 4% PFA and sectioned on a cryostat, using the MEC13.3 antibody (BD Bioscience) which I understand is the best. I'm not getting very good staining though, and am wondering if the PFA fixation is interfering with antibody binding (they do not recommend this antibody for formalin fixed paraffin embedded tissues). Any suggestions?
Mike
Any recommendations for immunhistochemistry protocols for CD31 staining? I am currently trying it on tissues which have been fixed in 4% PFA and sectioned on a cryostat, using the MEC13.3 antibody (BD Bioscience) which I understand is the best. I'm not getting very good staining though, and am wondering if the PFA fixation is interfering with antibody binding (they do not recommend this antibody for formalin fixed paraffin embedded tissues). Any suggestions?
Mike
1. when you want to stain cryopreserved tissue sections you don´t have to fix them with formaldehyde
2. formaldehyde crosslinks your proteins (lysins) therefore you have to perform an intermediate so called antigen retrieval. otherwise your antibody is not recognizing epitope. it could be complex.
3. i would suggest: try it without fixation
our CD8, CD4 and F4/80 staining worked well on cryopreserved tissue sections
-moljul-