Using 2 polyclonals (one rabbit, one goat) in double staining? - (Oct/26/2008 )
A question for you all to ponder
Can anyone see any problems with using 2 polyclonal antibodies in double immunofluoresence?
One is raised in goat and one is raised in rabbit- so different species.
However, They are both raised against the CT of their respective proteins.
The proteins are members of the same family of proteins (but are against different proteins) and polyclonal antibodies are all that are available to me.
If I see co-localisation- will it be accurate and not some artifact of cross-reactivity?
Any thoughts gratefully received
I do this often so I'd imagine you're probably going to be OK doing both at once as the primary's are from different species. You just have to be sure that your secondary Ab's don't react with each other or are from the same species of your cell source.
I thought it should be ok- the secondaries are ok (donkey), just wanted to cover myself- I'm fairly certain I should see some co-localisation, but I needed to reassure myself I've covered everything.