Some researchers face a tough issue in immunoprecipitation (IP)or co-immunoprecipitation (co-IP): the primary antibody they use in IP or co-IP is the same as the one they 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 less than 30kDa, in order to avoid interference of the antibody light chain , we recommend using heavy chain specific secondary antibodies, such as :
|Product name||Product code||Conjugate||Details|
|IPKine HRP Goat Anti-Mouse IgG HCS||A25112||HRP||Click here|
|IPKine HRP Goat Anti-Rabbit IgG HCS||A25222||HRP||Click here|
Abbkine’s IPKine HRP AffiniPure Goat Anti-Mouse(Rabbit ) IgG Heavy Chain secondary antibody reacts with with Fc portion of mouse (rabbit) IgG heavy chain, and it doesn’t react with the Fab portion of mouse(rabbit) IgG. It's cross-reaction with bovine , horse and human serum proteins has been specialized minimized (anti-rabbit antibody only absorbed with human serum protein) , while it may cross-react with immunoglobulins from other species. It has no reactivity on non-immunoglobulin serum proteins. Furthermore, they are conjugated to horseradish peroxidase (HRP), ideal for western blotting.
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