Enhanced chemiluminescence (ECL) for routine immunoblotting: An inexpensive alternative to commercially available kits.

Spermatogenesis (2012-02-10)
Dolores D Mruk, C Yan Cheng

Immunoblotting is an analytical technique used by many laboratories to study protein expression. It involves electrophoretic separation of proteins by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), immobilization of these proteins onto a membrane of either nitrocellulose or polyvinylidene difluoride, incubation of the membrane in a monoclonal or polyclonal antibody and detection by a standard method such as enhanced chemiluminescence (ECL). To achieve this, most laboratories opt to use commercially-available chemiluminescence kits which are acceptable but relatively expensive. In this technical report, we show that a self-prepared chemiluminescence reagent is superior to a commercially obtained kit in terms of sensitivity, duration of signal, ease-of-use and shelf-life but at a fraction of the cost of a kit.