Detection of chicken meat in raw meat mixtures by the double method of an enzyme immunoassay and an immunoblotting technique

Polyclonal antibodies were raised in male New Zealand White rabbits against a 0.5% (v/v) Lubrol PX protein extract of crude chicken red bone marrow. Following serum protein removal by ammonium sulphate precipitation, the serum was affinity‐purified against muscle protein from a selection of different species. The antiserum, when assayed by enzyme‐linked immunosorbent assay, was shown not to recognize any proteins from specimens other than chicken hand‐deboned (HDM) and mechanically separated (MSM) meats, with the reaction towards MSM chicken being significantly stronger than that against HDM chicken. When the same samples were separated by sodium dodecyl sulphate polyacrylamide gel electrophoresis, transferred to nitrocellulose by Western blotting and immunochemically stained with the affinity‐purified antiserum, they showed specific staining of protein bands within chicken HDM and MSM, with no reaction towards any proteins of any other species. A band common to both chicken HDM and MSM at about 45 kDa , ...