The preparation conditions will vary dependent on the antibody you are looking at, which may require some optimisation in terms of antigen retrieval steps or the fixation of the tissue. Can I ask if your slides were from tissue fixed (in say formalsaline) and wax embedded, or were cut as frozen sections?
A good starting point for fixed and wax embedded sections is to dewax with Zylene for 20 mins, followed by washing with graded ethanols (100%, 100%, 90%, 70%, 50% ) for 2 mins each and into water. An antigen retrieval of heating in citrate buffer (I use 0.1M at ph6) is probably the most commonly used, and the one I would try. Most of my antibodies work on skin tissue with citrate retrieval heating on full power in a microwave for 2-3 mins, and leaving to cool for a further 10 mins. Alternative retrieval methods using enzymes such as proteinase, EDTA or acids are possible. After the retrieval you follow the instructions for whatever kit you are using - I find the vectorstain ones are good. You would have to just try and see what works for your particular experiment.
Good luck!!
Hi David! Thanks for you answer!
My slides were from tissue fixed (in say formalsaline) and wax embedded.
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