questions about functional blocking antibodies

[wangleicau]

I am using a functional blocking antibody (mouse anti-human beta 1 integrin) to block the beta 1 integrin in S1 cells (Human mammary epithelial S1 HMT-3522 cells) before I do the immunostaining, but for the immunostaining I have no other choices but use mouse anti-human (NuMA) as primary antibody, and goat anti-mouse as the secondary. I am afraid that the secondary antibody I use will react with the functional blocking antibody. Will it happen? And is there anyone knows whether the functional blocking antibody I use will give me fake fluorescence? Can I wash the functional blocking antibody out?

Thanks.

[ethidiumbromide]

I'm not really sure, but I would tend to think that it would interfere. Is there any way you can remove the blocking antibody just prior to immunostaining? I'm sorry, I'm not familiar with the technique you are using. And is there any way you can get a rabbit anti-human?