Peroxidase labeling of IgMs fragment of ABO blood group specific mouse monoclonal IgM.
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.
Yukawa
N
N
Department of Legal Medicine, Miyazaki Medical College, Japan.
Matsuda
H
H
Seo
Y
Y
Suetomi
K
K
Takahama
K
K
D000017:ABO Blood-Group System / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D004355:Drug Stability; D006735:Horseradish Peroxidase / Q000737:chemistry; D006801:Humans; D015151:Immunoblotting; D018796:Immunoconjugates / Q000737:chemistry / Q000302:isolation & purification; D007124:Immunoenzyme Techniques; D007075:Immunoglobulin M / Q000737:chemistry*; D066298:In Vitro Techniques; D008301:Maleimides / Q000737:chemistry; D051379:Mice; D011487:Protein Conformation
D000017:ABO Blood-Group System; D000911:Antibodies, Monoclonal; D018796:Immunoconjugates; D007075:Immunoglobulin M; D008301:Maleimides; C043592:maleimide; D006735:Horseradish Peroxidase