During the fixation step of immunocytochemistry/immunofluorescence, some protocols say that cells can be stored at -20 for a week, while some protocols advice to keep it for 10 minutes to preserve the structure and epitope of the protein of interest.
What is the ideal time that we should use for better cell fixation without altering the epitope of our protein of interest?
And if over incubation in methanol does alter the epitope of protein, are there any antigen retrieval methods for such type of epitope alteration?
Dear Vikas, You have not clarified the cell types origin, however, cells fixation to methanol is enough for 10-20 mins at -20 degree.
Best,
Methanol already at -20C, 10 minutes to 20 minutes best. More time will not make any additional benefit.
Dear I support the idea of 10-20 minutes and proceed to staining without delay.
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