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Multiple Antigen Labeling

The preferred method for double or triple labeling involves sequential staining of each primary antibody. With proper color development of substrate, the reaction product from the first substrate usually prevents most of the subsequent antibodies and detection reagent(s) from interacting with components used to stain the first antigen. This feature allows multiple labeling with primary antibodies raised in the same species using a single VECTASTAIN® ABC kit or ImmPRESS™ reagent. The order of the primary antibodies and the substrates is very important. Several controls and additional blocking steps may be necessary to obtain optimal results. Numerous combinations of colors are possible using Vector Laboratories’ products and our multiple labeling protocols