TE antigen retrieval and immunostaining
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=== Principle === | === Principle === | ||
| + | |||
| + | * aldehyde treatment ([[Polymethanal (paraformaldehyde)|paraformaldehyde]], glutaraldehyde) chemically fixes cells and tissue | ||
| + | * [[Paraffin embedding and sectioning|paraffin-embedding]] allows very thin tissue sections to be cut | ||
| + | * however, some antibodies don't work well after aldehyde fixation and paraffin embedding as opposed to other fixatives and cryosections | ||
| + | * [[antigen retrieval]] methods can improve antibody binding by reversing chemical modification of epitopes or by unfolding/refolding epitopes | ||
=== Material === | === Material === | ||
Revision as of 14:49, 2 November 2009
This 2-part protocol describes antigen retrieval with Tris EDTA buffer followed by immunostaining. Antigen retrieval can be helpful for aldehyde-fixed, paraffin-embedded samples that don't give sufficient signal with a standard immunostaining protocol. Several antigen retrieval methods exist but, unfortunately, you need to determine empirically which one is most suited to your antibody.
Contents |
Deparaffinisation
Tris EDTA heat-based antigen retrieval
Principle
- aldehyde treatment (paraformaldehyde, glutaraldehyde) chemically fixes cells and tissue
- paraffin-embedding allows very thin tissue sections to be cut
- however, some antibodies don't work well after aldehyde fixation and paraffin embedding as opposed to other fixatives and cryosections
- antigen retrieval methods can improve antibody binding by reversing chemical modification of epitopes or by unfolding/refolding epitopes
