B-mercaptoethanol (BME) protocol?

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Wolfgang H. Muss

Buon giorno,

an interesting question/ request. Unfortunately I can't help you practically...but it will - hopefully - turn out ASAP who can provide an optimal protocol for using BME-ab-stripping solution in IHC/ImmFluor-staining methods. Meanwhile,

dear Daniele Pantano , just to forward proposals for initial readings (not knowing your - for sure - sufficient level of knowledge which I would like to attain myself too):

cf:

https://en.wikipedia.org/wiki/2-Mercaptoethanol
https://www.amherst.edu/system/files/Beta%2520Mercaptoethanol.pdf
http://bridgeslab.sph.umich.edu/protocols/index.php/SOP_-_B-mercaptoethanol
https://www.biorxiv.org/content/10.1101/533547v4.full.pdf (Manuscr. Ehrenberg et al, 2019; search for: BME-stripping solution, chapter 2.3.2 Cycle of staining and stripping - p.-9-)
https://www.sigmaaldrich.com/technical-documents/protocols/biology/western-blotting/buffers-recipes/detergent-stripping-buffer.html

Awaiting many answers of really skilled colleagues, and

Best of luck, Wolfgang

Patrick Lewis

I will also be doing an IHC for multiple markers on frozen samples and I am concerned about tissue fragility, and cell surface marker fragility. My advice such that it is trial and error and if that fails see if there are other pathways such as paraffin or better markers to use. Poor advice I know, but all I have right now.