I need a standardized protocol for Biotinylated WFA (Wisteria Floribunda Agglutinin) staining on PFA fixed mice brain sections.
Dear Idris,
Usually you can find suggestions how to use antibody or lectin on the websaite of the seller. I have looked for biotynylated Wisteria agglutynin from well known sources, Sigma and Vector laboratories both have it. I often use products from Vector labs, and they have a general protocol for IHC with biotynylated agglutinins - see attached link: http://docs.vectorlabs.com/protocols/Detect.Glycoproteins.pdf. The file is for general guidance - our final protocol will depend on your test experiments, you may have to adjust the concentration of your agglutinin depending on the length of fixation and what method you choose to detect biotin
Wolfgang Hartig papers list WFA lectin from Sigma, and quote concentration - but you still have to test as this work was in1990s: (good microphotographs) - an example:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1167753/pdf/janat00124-0105.pdf
Good luck with your experiments.
Thanks once more, Maria. I did look into your enclosed links. Tried out the protocols and the staining worked really nice.
Hi Idris, similar to Dominic, I am interested in staining for WFA. Any protocol you could provide would be greatly appreciated.
Hi Dominic and Phillip,
Enclosed here is my working protocol on WFA staining for PFA fixed, frozen mouse brain sections.
Let me know if you need further assistance/ clarifications.
Regards.
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