Dear all,
I have mouse brain tissue perfused with PFA 4% and post fixed in formol 10% for periods between 2 month and 1 year. I cut them on vibratome (30um) and I made inmunohistochemistry and beta galactosidase to check cellular senescence.
My problem is that I have less signal on positive controls if postfixation on formol was longer. There is a way to solve this problem?
Thanks in advance for your reply
I have used the microwave antigen recovery method, but on paraffin embedded sections. You may try on vibratome sections. The method is described in the enclosed publication in case you want to give it a try. I have used for antigen recovery on a variety of tissues, from different species and to recover a variety of antigens.
Dear Mariela,
Microwaving will definitely help! We use it on vibrotome sections and even on free -floating frozen sections!
Good luck!
Natalia
Dear Mariela,
Here you will found two protocols to atigen retrieval.
http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol
Good luck!
Dear Mariela,
Retrieval antigen step using microwave and citrate buffer.
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