A given laboratory may aim to detect different proteins of varied expression levels, in the tissue being analyzed. Agrisera offers a unique pack of combined reagents, with two different sensitivities. This allows more versatile detection, of high and medium abundancy proteins (Agrisera ECLBright), as well as low abundancy proteins (AgriseraECL SuperBright). Depending on the target protein that we aim to detect, different detection reagent needs to be used, and Agrisera offers this possibility provided in the same set.
In the example above, the aim was to detect a highly abundant photosynthetic protein, PsbA. Using Agrisera anti-PsbA antibodies (AS05 084), a matching secondary antibodies of high titer, goat anti-rabbit, HRP-conjugated (AS09 602) and Agrisera ECL reagent of extreme low femtogram detection range (AgriseraECLSuperBright), resulted in prominently increased background signal. Choosing the other reagent, provided in the Agrisera ECL set, gave the possibility to optimize the result in a very swift manner. There is no need to repeat the whole procedure, as the membrane can be subjected to a wash and developed with less sensitive ECL. It is oftentimes not the primary antibody that contributes to increased background, but rather high load/well, combined with a too sensitive detection reagent.
We are always keen to discuss your results and advice promptly. Contact Agrisera support either using a chat button on the website or by email to [email protected].