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|- Incase of Low CYTO-ID Green dye staining in all treatments, increase the reagent concentration (500X dilution of the dye is recommended) and the incubation time.
- Incase of High CYTO-ID Green staining, it could be that the cell culture medium was depleted of nutrients and so change media 4-8 hours before the experiment.
|- incubate primary Ab with agitation for 1 hour.|
|- If high background or Non-specific bands is present,Try different blocking strategies: Longer blocking times, higher % of blocker, a different blocker, inclusion of blocker protein in antibody dilutions or try a fresh batch of the same blocker.|
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