Hello,
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?
Best,
Matt
Inter cellular stain on mouse frozen tissue
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mattbliss90
- Posts: 10
- Joined: Wed Jan 17, 2018 11:37 pm
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ssivajothi
- Posts: 32
- Joined: Thu Jan 11, 2018 9:00 pm
Re: Inter cellular stain on mouse frozen tissue
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.
Santhosh
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.
Santhosh
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MariekeIJsselsteijn
- Posts: 8
- Joined: Mon Jan 15, 2018 10:52 am
Re: Inter cellular stain on mouse frozen tissue
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.
Marieke
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.
Marieke