Figure 3
From: Mouse Antibody of IgM Class is Prone to Non-Enzymatic Cleavage between CH1 and CH2 Domains
Legumain does not cleave mouse IgM. (a) IgM cleavage in serum treated with alkylating agents. Alkylating agents, such as iodoacetamide or NEM, are potent inhibitors of legumain – a cysteine protease specific to Asn at P1 site. Activity of legumain present in FBS was tested using fluorogenic substrate Z-Ala-Ala-Asn-AMC. Samples were analyzed using western blotting with anti-mouse IgMκ antibody. The full-length blot is presented in Supplementary Figure S2. (b) Recombinant legumain did not cleave IgMs. The gel was stained with Coomassie BB. (a and b) Representatives of two independent experiments are shown.
