Has anyone used the Subcellular Protein Fractionation Kit for Cultured Cells by ThermoScientific?
I am very confused about the last fraction in this kit, which indicated as "cytoskeletal". But when I blotted my samples with A-tubulin or actin, most of the signal signified at cytosolic fraction, with nothing in the "Cytoskeletal" fraction at all.
While there is lamin A signal at the "Cytoskeletal" fraction. Does it actually means the nuclear skeleton?
Many many thanks!!
I have a similar confusion experience on this fractionation work with my homemade buffer as well. I think the main cause for this controversy is from the suboptimal buffer which released unscheduled proteins from the cells. If possible, try some immunofluorescence work may avoid this confusion.
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