F-actin label in LIVE cells
gal-1 at bones
Fri Feb 17 09:33:28 EST 1995
On 15 Feb 1995, Tobias Baskin wrote:
> This may be "stating the obvious" but for many years folks have
> been labeling actin in cells with rhodamine phalloidin. There are a number
> of variants, different fluorophores etc, Molecular Probes will have lots of
> them, and info/citations in their catalog. While it is true that this
> compound can exert a stabilizing effect on actin filaments, it is typically
> used as a label at concentrations well below those where obvious
> stabilization effects occur.
No-one uses phalloidin for live cells, it is essentially impermeable
(except for hepatocytes). Most of the live actin staining that I have
seen has been done by loading the cells with fluorescently labelled actin.
Although but G actin and F actin will be labelled, it turns out that the
intensity of the F actin is far and away the largest signal. Try looking
at some of the work from D. Lansing Taylor or Marc Kirschner to get the
details and see if it will work for you.
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