[Histonet] Lectin IHC
Hi To all,
Here is more detailed descripiton of what I have done. The organism I
am studying is Guinea Pig. The tissues are infiltrated with OCT and kept
frozen until cut.
This is the General Protocol of my lectin staining
1. Air Dry for 15 mins
2. Application of fixative (depending on the reagents used, incubation
time changes) and tap water rinse
3. Block for 30mins (3X Buffer Wash)
4. Lectin Incubation for 1 hr (3X Buffer Wash)
5.Application of ABC for 30 mins(3X Buffer Wash)
6. Application of substrage for 15 mins (tap water rinse)
7. Counterstain with mayer's hemotoxylin
The combinations that I have tried are ( fixitive/block/dectection
10% Formalin/ universal block/ avidin-biotin
10% formalin/ 0.1% BSA block/avidin-HRP or avidin-Biotin
Bouin/universal block or 0.1% BSA block/avidin-biotin
I haven't been able to get a positive staining for my positive control.
The lectin I been using is UEA-1 Any suggestion would be great.
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