Immunofluorescence (IF) is a method used to visualize a protein’s distribution in a sample (fixed cells or tissue), viewed under a fluorescent microscope. This method relies on fluorescently conjugated antibodies that bind to an epitope of a target. The fluorophore may be conjugated to the primary antibody for direct IF or it may be conjugated to a secondary antibody for indirect IF.
Although direct IF is more convenient, not all primary antibodies are commercially available as fluorophore conjugates. To overcome this obstacle, Advansta offers an array of antibody conjugation kits to suit your detection needs. Our SpectraDye antibody labeling kits require as little as 10µg of purified antibody, the reaction is complete in 30 minutes and there is no need to purify the conjugate after the labeling reaction is complete. If you are pressed for time or have two or more primary antibodies produced in the same species, this is definitely the way to go.
For detection of very low abundance targets it may be advantageous to stick to an indirect IF protocol. Like any method, this one also has a few disadvantages. Some major issues with indirect IF detection are high general background, non-specificity and cross-reactivity of secondary antibodies. To address these issues, Advansta offers SpectraDye secondary antibodies. These conjugates produce bright signal with high specificity since they are pre-adsorbed against irrelevant serum proteins.
Regardless of which IF method you choose, Advansta’s SpectraDye antibody labeling kits and secondary antibodies will help you achieve bright, clean, publication quality results.
SpectraDye Secondary Antibodies
SpectraDye Antibdoy Labeling Kit
AdvanBlock-Fluor Blocking Solution