I'm working in the interaction ALCAM-ALCAM in metastasis models. I wondered if anyone checked the basal expression of ALCAM in B16F0 and/or B16F10. I have observed different results of ALCAM level expressions by FACs in different experiments and I don´t know what is happening. I used the same conditions in all the replicates. On the other hand, do you recommend me a western-blot to analyze that?
Thank you so much for your reply!
I never used ALCAM on B16 but I extensively stained human culture with ALCAM. quite often they show heterogeneous expression, so WB should mask this heterogeneity that could be crucial in metastasis model (only few cells are really metastatic, or have unlimited metastatic potential). I always reccomand FACS to study populations, ALCAM can be modulated by different cellular conditions, I mean different cell cycle phases or different proliferation rate, make sure your cells are always cultured at same density and you use them after same time after seeding. good luck!
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