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Peroxidase labeling of IgMs fragment of ABO blood group specific mouse monoclonal IgM

N Yukawa1, H Matsuda, Y Seo

  • 1Department of Legal Medicine, Miyazaki Medical College, Japan.

A method for peroxidase labeling of the monomeric subunit (IgMs) of ABO blood group specific mouse monoclonal IgM is described. IgM was purified from a commercial monoclonal anti-B blood grouping reagent by a combination of salt precipitation, euglobulin precipitation, and gel filtration. IgM was mildly reduced with L-cysteine to yield SH-bearing IgMs. Finally, IgMs was conjugated to horseradish peroxidase, into which SH-reacting maleimide groups had been introduced using N-succinimidyl 6-maleimidohexanoate, through the selective reaction between SH of IgMs and maleimide groups of peroxidase.

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