How to choose secondary antibodies?

The secondary antibody is selected based on the source of the primary antibody and the customer's experimental needs. First look at the species origin and subtype of the primary antibody used in the experiment. Determine the secondary antibody based on the species source and subtype of the primary antibody: if the primary antibody is rabbit-derived IgG, then choose an anti-rabbit IgG secondary antibody, such as Goat Anti-Rabbit IgG; if the primary antibody is mouse-derived IgM, then choose Choose an anti-mouse IgM secondary antibody, such as Goat Anti-Mouse IgM. Marker selection. Common markers include HRP, Biotin, fluorescein, etc. ELISA commonly uses Biotin-labeled secondary antibodies, followed by a streptavidin-labeled HRP for further signal amplification, and TMB substrate for color development; WB mainly selects HRP-labeled secondary antibodies and uses ECL substrate luminescent solution for color development; IHC generally chooses HRP-labeled secondary antibodies. It is recommended to use an immunohistochemistry kit (E-IR-R217 or the latest E-IR-R220/221), and generally use DAB substrate solution for color development; IF can choose secondary antibodies labeled with different fluoresceins according to experimental needs, such as FITC, Cy3, PE, etc. (try to avoid the autofluorescence of cells), and detect the fluorescence through a fluorescence microscope.