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Hello,

I have a question about blocking the culture in immunofluorescence staining which is done prior to the addition of the stain-bound secondary antibody. I do not understand what blocking does. In one reference I read it that adding the blocking solution actually blocks the large plasma proteins that may cause background staining. Another reference said that blocking is done to block those IgG antibodies that may be present in the culture and are against the animal in which the secondary antibody was raised. I'm really confused.

shadirjb wrote:Hello,

I have a question about blocking the culture in immunofluorescence staining which is done prior to the addition of the stain-bound secondary antibody. I do not understand what blocking does. In one reference I read it that adding the blocking solution actually blocks the large plasma proteins that may cause background staining. Another reference said that blocking is done to block those IgG antibodies that may be present in the culture and are against the animal in which the secondary antibody was raised. I'm really confused.

Hello, shadirjb!

I think that your question is fascinating, but I also think that you would be more likely to get help by posting your question to the Life, Earth, and Social Sciences forum. The experts that participate in that forum have expertise in biology and other sciences that are better suited to your question.

Best wishes for a successful project!