What should I consider when choosing antibodies for my western blot experiment?

There are several things to consider when choosing antibodies for western blot experiments. One thing to consider is that the primary antibody should exclusively recognize the protein of interest without cross-reacting with other proteins present in the sample. One should also use an antibody targeted against a conserved region of the protein to enhance its compatibility across different species. Another thing to consider is to use an antibody that has been validated for western blotting, which confirms its effectiveness in producing a strong signal. Additionally, one should select a primary antibody raised in a different species than the sample's organism to prevent cross-reactivity with endogenous immunoglobulins. Additionally, the antibody should have a high affinity for the target protein, ensuring tight binding even at low concentrations. Another consideration is to ensure that the primary antibody's isotype is compatible with the secondary antibody to be used. Lastly, one should determine the optimal concentration of the primary antibody through titration protocols. Using too much antibody may result in background noise, while using too little may yield weak signals.