How does R & D Antibodies perform ICC experiments on cells and tissues?
Initially, one should perform a titration experiment to optimize the signal to noise ratio especially if peptide or protein blocking experiments are to be conducted. However, we have successfully used our monoclonal antibodies as culture supernatants, our monoclonal antibodies as ascites fluids, and our polyclonal antibodies at dilutions of at least1:50, 1:500 and 1:1000, respectively (see specific data sheet for individual antibody). Dilute the stock antibody in PBS, pH 7.2, containing either 0.1% BSA or 1% normal goat serum. For cells cultured on slides or coverslips We wash the cells 4 times in PBS pH 7.2, rinse once quickly in distilled water, and drain well. We fix the cells with either 70% ice cold acetone or neutral buffered formalin for 10 min, air dry and store them frozen. We always allow the slides to warm to room temperature before using. If the cells were fixed with organic solvents, such as 70% acetone, 95% ethanol, or methanol, then they do not need permeabilizing. Howev
