Animal handling: While planning a c-Fos experiment with animals you should be aware of technical pitfalls affecting your research. For instance, you should consider that c-Fos expression is time dependent, susceptible for stress and can be detected after minutes (Herrera and Robertson, 1996). It is important to take into account that many factors like animal handling itself (e.g. needle injection) or the exposure to light during the dark phase (circadian activation) can influence c-Fos expression (Asanuma and Ogawa, 1994; Herrera and Robertson, 1996). Therefore, the experiment has to be planned and conducted accurately with appropriate control groups to avoid false positive results.
IHC protocol: Another point you should care about is your immunohistochemical (IHC) protocol. It seems that the total number of c-Fos positive cells is threshold dependent. Many parameters affect the detectable number of c-Fos positive cells: tissue storage conditions, method of sectioning (cryostat or vibratome), staining parameters (e.g. incubation temperature), signal enhancing reagents (Berghorn et al., 1994; Werner et al., 1996; Mayer and Bendayan, 2001) and, above all, primary antibody selection.
c-Fos antibody: Primary antibodies differ in their specificity, sensitivity and signal to noise ratio and these properties affect the outcome of your experiment. Therefore, we aim to provide highly specific and consistent c-Fos antibodies to support scientific research and experimental reproducibility. Since monoclonal antibodies have several advantages compared to polyclonal antibodies, we developed a new monoclonal recombinant rabbit anti-c-Fos antibody.