Incubation Time of Primary Antibody

The concentration of the primary antibody, incubation time and temperature all will affect the quality of the immunohistochemical staining, these variables need to be optimized for different antibodies, in order to achieve high specificity and low background immunohistochemical staining, the optimization process is generally to maintain a fixed incubation time and temperature, while varying the concentration of the antibody, for example, the use of high affinity antibodies, may be used a relatively high concentration of antibody with incubation time shortened, or a relatively low concentration antibody with longer incubation time, in order to obtain highly specific staining, often choose a longer incubation time and lower incubation temperatures.