anti-flag antibodies - (Aug/07/2007 )

Hi everybody,
I need to know if it is possible to use anti-flag antibodies (such as anti-his, anti-myc or others) to label membrane proteins in living cells in a culture.
I'd like to know which one, and from what company, binds its target epitope with the highest avidity and specificity.
Thanks

The antibodies that you mentioned above (anti-flag, HA, Myc...) are all usually raised against a peptide of the epitope. Thus they will be able to recognize that special peptide. When you use fusion protein with these tags, the antibodies will be able to recognize them (unless your tag is masked somehow, and that is rare IMHO). It does not matter if the tag is outside or inside the cells, just that if the tag is inside, then living cell staining is unsuitable. Of course, sometimes there could be background if other proteins in cells also got the particular peptide, but in most cases the problem is not serious and you can easily discern it using proper controls.

We have used the antibodies from a few companies and they are all fine. Sigma and Upstate are two good sources.

Hi,

I usually use the mouse-anti-FLAG M2 from Sigma without having problems... but it's important to dilute this antibody quite well...

Chakchel