HI,

my having trouble with my positive staining on paraffin tissue with immunofluorescence..

The exposure time required is very high since the signal is soo dim... it takes a long time to take pics...
I'm not sure what the problem could be?.
I would really appreciate any help?

Can you increase the concentration of your primary antibody (i.e. lower dilution of your antibody) to make the signal stronger?

Hi Tracy,

I'm just worried if increase the concentration if that would cause too much non specific staining but I can try it..
thanks

I agree with your concern about increasing the antibody concentration. this usually just makes antibody stick in places it shouldn't.

What fluor are you using? Labeled primary or secondary? Homemade or commercial?

I am using a Donkey anti RabbitIgG conjugated to Cy3- a labelled secondary