[Histonet] Tips on staining PLA2R assay on Leica Bond

[Eddie Martin]

I don’t do staining for this antibody, but have experience with IMF on frozen tissue and FFPE and have worked with the Leica BOND max and BOND-3.  If you would prefer IHC over IMF, you can search for a human anti-mouse, or human anti-rabbit antibody.  Abcam provides a polyclonal whole serum antibody that works on FFPE.  I grabbed this from their website: 
Anti-PLA2R antibody (ab80054)

Since Abcam’s PLA2R antibody is a rabbit anti-human, you would need to create a modified DAB or modified RED protocol that doesn’t include the secondary antibody / linker step in your protocol prior to dispensing polymer.  The rest of your protocol can remain as it is.  

As you are using a Leica BOND, both ways are actually pretty easy to set up, either indirect IMF or using Leica’s DAB kit for IHC. You would just need to play around with retrieval times with both Citrate and EDTA to get the optimum staining pattern. Additional ancillaries may be added when necessary should you have any background stain that is hard to get out.

Best Regards,

Eddie Martin, HT(ASCP), QIHC
954-826-9403
Edmartin26 at gmail.com