IF with hybridoma supernatant - (May/28/2010 )

Dear colleages,

does anybody have experience in doing immunofluorescence with a primary antibody from hybridoma supernatant? Do you have background problems?

Best,

Daniel

I've done IF with CHO, HeLa and U2OS cells using the hybridoma supernatant for the anti-HA (clone 16B12). I blocked the cells as normal and it worked just as well as the purified antibody. I just added 1mL of the supernatent to 9mL of BSA and stained as usual (one hour, room temp). You may need to work out the conditions that work best for your specific antibody.

Okay, I'll try. Thanks alot!