Could anyone help me with staining T cells for surface activation markers (CD69) and Ki67/Granzyme B at the same time? The Ki67 staining protocol requires 70% ethanol treatment which might be too harsh for the other markers staining? Thanks!
Dear Jingjing Shen
Check out my rather detailed answer to a similar type of question from 2 years ago.
https://www.researchgate.net/post/Problems_with_Ki67_staining_to_look_at_T_cell_activation_proliferation
The details concerning the reagents (antibodies, clones and flourophores) as well as the experimental protocols you'll find in those two papers.
Article Deubiquitinase MYSM1 Is Essential for Normal Fetal Liver Hem...
Article A role for the histone H2A deubiquitinase MYSM1 in maintenan...
I hope that helps. Please let me know, if you have any additional questions.
All the best and good luck with your experiments,
Michael
Dear Jingjing Shen,
You can find the answer in this nature published paper PMID: 30089911
Best regards
We published a protocol for sequential staining of a single slide. This allows combination of your Ki67 protocol with the other staining (EtOH step in the final stage).
Zuiderwijk M, Geerts M, van Rhijn CJ, van den Bogaerdt A, Hamming JF, van Dijk RA, Lindeman JH. Leukocyte Dynamics during the Evolution of Human Coronary Atherosclerosis: Conclusions from a Sevenfold, Chromogen-Based, Immunohistochemical Evaluation. Am J Pathol. 2018 Jul;188(7):1524-1529.
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