How do I control for specific (immuno) staining in my experiment ?
It is key to any immunofluorescence experiment that the staining of cell structures or proteins is specific. Controls for specific staining are categorized for antibody specificity and method specificity. All the followig controls should be consifdered carefully, especially when trying a new antibody or a known antibody on a new preparation. Controls for antibody specificity i.e. does the antibody recognize the target protein specifically. Be aware that under certain circumstances (e.g. particular fixation protocols) antibodies can bind highly selective to a non-specific epitope (see Willingham,1999 for details). • Preabsorbtion of antibody with the protein/peptide that was used to generate the antibody before incubation of the specimen. This should block the specific binfing sites of the antibody and reveal non-specific binfing of the antibody in the tissue or cells • Westernblot/Immunoblotting shows that the antibody recognizes specifically a protein of the correct size • Immunopreci
