This is not for microscopy studies. This is for an in cell protein foot printing technique, Fast Photochemical Oxidation of Proteins FPOP.
I just need guidance on how to get the unlabeled phalloidin and Cytochalasin B into live cultured HEK cells (Plate size, Concentrations, Incubation time, other parameters to consider?)
I want to harvest the cells for the IC-FPOP experiment, then digest to run on Mass Spec.
I appreciate any and all feedback.
Thank you!
You don't need to worry about the Cyto B. It penetrates the plasma membrane within a few minutes. Phalloidin does not, so you need a carrier for that.
Hi Dante, both compounds are cytotoxic and will kill most cells. So you will have to use concentrations that are sub-toxic or you can go for broke and incubate the cells with a lethal dose.
All cell lines have some heterogeneity and you might be lucky to select for a clone that is resistant to lethal doses of these compounds.
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