How should I choose the secondary antibody?

How should I choose the secondary antibody?

The secondary antibody is selected according to the source of the primary antibody and the customer's experimental needs. Firstly, we will look at the source and subtype of primary antibody used in the experiment. Decide the secondary antibody according to the source and subtype of the primary antibody: if the primary antibody is rabbit IgG, then choose the secondary antibody against rabbit IgG, for example, Goat Anti-Rabbit IgG; if the primary antibody is mouse IgM, then choose the secondary antibody against mouse IgM, for example, Goat Anti-Mouse IgM. Selection of markers. Common markers include HRP, Biotin, fluorescein, etc. Biotin is commonly used in ELISA. ELISA commonly used Biotin-labeled secondary antibody, followed by a streptavidin-labeled HRP to play a further signal amplification, with TMB substrate color development; WB mainly choose HRP-labeled secondary antibody, with ECL substrate luminescent liquid color development; IHC is generally also the choice of HRP-labeled secondary antibody, recommended to use immunohistochemistry kits, generally with DAB substrate liquid color development; IF according to the needs of the experiment to choose a different fluorescein labeled secondary antibody. IF can choose different fluorescein-labeled secondary antibody according to the needs of the experiment, such as FITC, Cy3, PE, etc. (try to avoid the spontaneous fluorescence of the cells), and detect fluorescence by fluorescence microscope.