Inter cellular stain on mouse frozen tissue

Does this antibody work? Designing a new panel?
Antibodies, isotopes, etc. for Imaging Mass Cytometry applications
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Inter cellular stain on mouse frozen tissue

Post by mattbliss90 » Tue Nov 13, 2018 7:00 pm

Has anyone here ever tried doing an inter-cellular stain on frozen mouse tissue? If so what protocol do you use... do you do a methanol permeabilization before adding all the surface antibodies and IC antibodies, or split the surface antibody incubation and IC antibody incubation into two different steps similar to what you would do for cells in suspension mode?

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Re: Inter cellular stain on mouse frozen tissue

Post by ssivajothi » Thu Dec 13, 2018 4:54 pm

Hi Matt,

I am wondering if you found any solution to this. Is a separate permeabilization step even required? In my trials with human frozen tissue, Ki-67 worked pretty well without any change to the standard protocol, but that was the only non-surface antigen I looked for.

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Re: Inter cellular stain on mouse frozen tissue

Post by MariekeIJsselsteijn » Tue Aug 06, 2019 9:13 am

Dear Matt,

I assume you solved this months ago but in case anyone else wonders in the future.
In our experiments we do a 5 minute fixation with cold metanol directly after cutting and then use both surface and IC antibodies together. We have a number of IC markers and they all work properly.

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