Immunofluorescence Tips

Primary antibody incubation:

Being the most important step in immunohistochemistry protocol, the primary antibody incubation is affected by incubation time and antibody concentration. As for the incubation temperature, there are 3 options: 4°C, room temperature and 37°C. 4°C is proved to be the best incubation temperature among the three, and the corresponding incubation time is overnight, and better to be followed with a rewarming (to 37°C)step. For the incubation at 37°C, the incubation time is normally 1-2 hours.