Myosin II Live Reporter

[Alison]

I would like to visualise myosin II while live imaging, without tagging myosin II with a flourophore. Is there a stain/dye that can be used for myosin II in live cells?

Thanks!

[Dagl1]

37 minutes ago, Alison said:

I would like to visualise myosin II while live imaging, without tagging myosin II with a flourophore. Is there a stain/dye that can be used for myosin II in live cells?

Thanks!

Question, when you say "tagging" with a fluorophore, do you mean things like FLAG or HISBIO tags or do you mean; "I don't want to anything to bind to myosin"?
You could just use a regular antibody and a secondary with fluorescent or histochemical activity (although I am not sure if IHC can be used for live cell imaging). 

If you mean you do not want anything to be bound to it (possibly because it may inhibit the movement of myosin?) you could also see if there is any other protein bound to myosin which you could instead target with antibodies.

-Dagl

[Alison]

25 minutes ago, Dagl1 said:

Question, when you say "tagging" with a fluorophore, do you mean things like FLAG or HISBIO tags or do you mean; "I don't want to anything to bind to myosin"?
You could just use a regular antibody and a secondary with fluorescent or histochemical activity (although I am not sure if IHC can be used for live cell imaging). 

If you mean you do not want anything to be bound to it (possibly because it may inhibit the movement of myosin?) you could also see if there is any other protein bound to myosin which you could instead target with antibodies.

-Dagl

Thanks! But I believe it is not possible to use IHC for live cell imaging.

[Dagl1]

2 hours ago, Alison said:

Thanks! But I believe it is not possible to use IHC for live cell imaging.

But using antibodies and a secondary which contains a fluorescent probe works for your experiment or do you want to specifically not use anything that directly binds it?

[CharonY]

3 hours ago, Alison said:

Thanks! But I believe it is not possible to use IHC for live cell imaging.

For live cell imaging tag-based covalent (self-) labeling is quite useful. There are range out there, including SNAP and  Clip.

[Prometheus]

You could look into Raman spectroscopy, doesn't require any tagging/staining. It comes with it's own problems, but might suit your needs.