In this study it turned out that only the labelling of antibodies using p-SCN-Bn-DOTA (2-(4-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) lanthanide chelates could be applied successfully in the Western blot approach whereas iodine labelling of the antibody showed unspecific binding and a too low sensitivity. After the immuno-reaction of the labelled antibodies with the antigen the element label is detected by laser ablation ICP-MS directly on the Western blot membrane. For this purpose target protein standards (bovine serum albumin, chicken lysozyme and bovine beta-casein) were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and electroblotted onto NC membranes. ![]() The labelling of three different polyclonal antibodies (chicken anti-bovine serum albumin, rabbit anti-chicken lysozyme and rabbit anti-bovine casein) with iodine and lanthanides has been investigated and the reaction conditions were optimized for application in a Western blot assay.
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