How can I combine a primary antibody with an immunofluorescence secondary antibody?

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Florian Brettner

Hi Dirk,

yes, thats right, the first antibody is an anti CD138-(goat IgG) and the secondary antibody is an anti goat IgG conjugated with Alexa Fluor546nm...

Thanks for the moment!

Florian

Korcan Ayata

Why do you want to link them before?

How will you remove un-bound secondary antibodies before you do incubation with the tissue?

You better use an antibody labeling kit if you have background florescence issues due to the secondary antibody.

Many secondary antibody that are bound to the primary antibody will result in a big complex that could weaken specific binding of the primary antibody due to changes in the charge. It will be worse If your primary antibody is also polyclonal.