I have encountered a significant discrepancy between my Fluorescein Phalloidin staining results and those reported in the literature. Typically, Fluorescein Phalloidin staining produces filamentous patterns; however, my results appear granular with unclear boundaries.
In my experiments, I stained HuH7 and Hep3B cells at a dilution of 1:1000 and fixed them with 4% formaldehyde for 20 and 60 minutes, respectively. Despite varying the fixation time, the results remain consistent.
Could anyone provide insights into potential reasons for this difference? Are there specific factors or troubleshooting tips that could help clarify these staining outcomes? Your expertise would be greatly appreciated!
Hi Marine, results may vary across different cell types. For our product validation, we used HeLa cells. Differences between cell types may lead to situations like the one you observed, so it may be helpful to check related experiments and literature on different cell types for reference. This article (PMID: 29470830) contains images showing good staining results—you could refer to its protocol and make slight adjustments to your steps.
We also found another paper that used phalloidin labelling in HuH7 cells, and the staining outcome looks quite similar to your case (PMID: 29163752).
The answer to this question is provided by MedChemExpress Technical Support.
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