title: Bead-based custom synovial antigen array_Stanford_Robinson Lab_v1.0
status: Public on Sep 10 2011
contact.city: Stanford
contact.first: William
contact.last: Robinson
contact.email: wrobins@stanford.edu
contact.state: CA
contact.address: 269 Campus Dr
contact.country: USA
contact.institute: Stanford University
contact.phone: 6508491207
organism: Homo sapiens
description: Bead-based multiplex immunoassay
  
  Fibrinogen A was purified from human blood, vimentin was purified from bovine lens, and histone 2B was purified from calf thymus.  All peptides represent human sequences. Anti-human IgG-PE secondary antibody was a goat monoclonal antibody.
  
  A custom made panel of putative rheumatoid arthritis associated autoantigens consisting of native and/or citrullinated proteins and peptides were conjugated to Luminex beads (Bio-Plex, Bio-Rad Laboratories, Inc).
manufacture_protocol: Peptide antigens were synthesized (by Fmoc chemistry) with N-terminal biotin. Bio-Plex beads were coupled to streptavidin using imide chemistry. The biotinylated peptides were then conjugated to the streptavidin-coated beads. Protein antigens were directly coupled to Bio-Plex beads using imide chemistry.