You may meet this problem in immunoprecipitation (IP) or co-immunoprecipitation (co-IP): the primary antibody you use in IP or co-IP is the same as the one you have used in WB, or they raised from the same host. When this happens, the secondary antibody will recognize the primary antibody used in WB as well as the denatured/reduced primary antibody released during the IP procedure . It directly result in two bands: the heavy chain (50kDa) and the light chain (25kDa).
If the size of your target protein is greater than 30kDa, in order to avoid interference of the antibody heavy chain , we recommend using light chain specific secondary antibodies, such as :
|product name||product code||Conjugate||Details|
|IPKine HRP AffiniPure Goat Anti-Mouse IgG Light Chain||A25012||HRP||Click here|
|IPKine HRP AffiniPure Mouse Anti-Rabbit IgG Light Chain||A25022||HRP||Click here|
Abbkine’s IPKine HRP AffiniPure Goat Anti-Mouse (Mouse Anti-Rabbit ) IgG Light Chain reacts with kappa light chain on mouse(rabbit) IgG. It doesn’t react with the heavy chain of mouse(rabbit) IgG. Their cross-reaction with human, rat, goat, sheep and bovine serum proteins has been specialized minimized, but they may cross-react with immunoglobulins from other species. They have no reactivity on non-immunoglobulin serum proteins. Furthermore, they are conjugated to horseradish peroxidase (HRP), ideal for western blotting.
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