We would like to see the protein expression for (IFIT3 gene which is nuclear protein) using Immunohistochemistry. Do I need to use Dapi-Fluoromount during the mounting steps. Here I have attached the protocol Can someone please review the protocol. Thanks
you may use or may not use. if you want Dapi, you can treat separately also. Be sure your nuclear secondary color channel is different from DAPI
Thank you so much Dr. Subhash C. Juneja for your answer. Can you please review my protocol for IHC. I would like to see the protein expression
Hello,
my advice is to have your tissue processed at a histology lab at your institution. Give them the tissue in formalin and have it processed and give you back the paraffin block.
In regards to the IHC method, you need to be clear to if you are using immunofluoresnce detection or light microscopy (immunohistochemistry). If you are using immunohistochemistry (IHC) then what detection system are you using: peroxidase or alkaline phosphatase? The choice of mounting really depends on the detection system. Also, no one can really give you detailed advice on a IHC protocol since these vary depending on what you re trying to accomplish, your antibody and your detection method.
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