[Histonet] Help with immunofluorescence
We are trying to demonstrate SHH and Survivin in human embrionic
colo-rectal tissue by Immunofluorescence in FFPE samples.
We are using:
* goat polyclonal SHH (N-19) sc-1194 with donkey anti-goat
* mouse monoclonal Survivin (D-8) sc-17779 with donkey anti-mouse
As antigen retrieval we have used saponin 0,1% in PBS in some slides=20
and a post-fixation in carnoy in others.
Carnoy postfixed slides seem to shoe a better image. However,and
specially in Sonic, we are seeing that some cells in the submucosa=20
appear labeled while epitelial cells in the mucosa remain unlabeled
As we are seing these images in samples treated only with the conjugated
Ab but also in samples with primary and conjugated, we think it could be
a cross or unespecific reaction of the secondary Ab or may be the=20
primary has not
May you help us at this point?
We would also ask you for some suggestion about tissue controls for=20
Any advice would be appreciated.
Thanks in advance
Tècnic Especialista en Anatomia Patològica i Citologia
Departament d'Anatomia i Embriologia Humana
Facultat de Medicina (HCP)
Universitat de Barcelona
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