The human sera containing anti-I and anti-i IgM autoantibodies anti-I Ma and anti-i P1A ELL, not described previously, and the mouse monoclonal antibody Fc10.2 were from the collection in Glycosciences Laboratory. The specificities of anti-I Ma and Fc10.2 had been assigned on the basis of inhibition of binding assays7 (link). Anti-I Ma binding was inhibited by the trisaccharide sequence Galβ1-4GlcNAcβ1-6Gal- on a branched backbone, whereas binding by mAb Fc10.2 was inhibited by the tetrasaccharide LNT: Galβ1-3GlcNAcβ1-3Galβ1-4Glc. Biotinylated anti-human IgM (μ-chain specific) and anti-mouse IgM were from Vector Laboratories (Cambridge, UK) and Sigma (Dorset, UK), respectively. Biotinylated RCA-120 was from Vector Laboratories.
Biotinylated rca 120
Manufactured by Vector Laboratories
Sourced in United States
Biotinylated RCA-120 is a reagent produced by Vector Laboratories. It is a lectin derived from Ricinus communis (castor bean) that has been labeled with biotin.
Automatically generated - may contain errors
2 protocols using biotinylated rca 120
1
Characterization of Anti-I and Anti-i Antibodies
2
Kinetic Assay and ELISA for C1-INH Activity
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PdC1-INH (Cetor®) was obtained from the Sanquin Blood Supply Foundation, The Netherlands. Monoclonal antibody (mAb) RII and polyclonal antibodies against C1-INH were produced previously [36 (link),37 (link)]. C1s for kinetic assays and ELISA was purchased from Calbiochem (La Jolla, CA, USA). C1s and pAb against C1-INH were biotinylated using EZ-Link® Sulfo-NHS-Biotin (Thermo Fisher Scientific, Waltham, MA, USA) or Pierce (Rockford, ILL) following the manufacturer protocol. Human Factor XIIa and kallikrein were purchased from Kordia (Leiden, The Netherlands). Agarose-bound RCA120 and biotinylated-RCA120 were from Vector Labs, USA. Streptavidin-peroxidase was obtained from Amersham Pharmacia Biotech (Uppsala, Sweden). Streptavidin-polymerized peroxidase (poly-HRP) was obtained from Sanquin (Business Unit Reagents, Amsterdam, The Netherlands). Chromogenic substrates S2251, S-2302 and S2366 were from DiaPharma (Beckett Ridge, Ohio, USA); C1s substrate H-D-Val-Ser-Arg.p-NA.2HCl was custom synthesized by Biomatik (Wilmington, DE, USA).
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