Bio-Rad ProteinChip Antibody Capture Kit - Manual (Page 2)

© 2007 Bio-Rad Laboratories, Inc.
Introduction
Traditional immunoassay-based protein detection methods are often difficult to
develop and are complicated by antibodies cross-reacting and binding
nonspecifically to a variety of proteins and peptides. The ProteinChip antibody (Ab)
capture kit enables unambiguous species-specific protein identification and epitope
discovery experiments on one platform, and eliminates the need to develop a
sandwich assay. Using the components provided in the kit, antibodies are bound to
a ProteinChip array to detect antigen(s) of interest from crude samples derived from
in vivo or in vitro experimental models. Once an antibody-antigen complex has been
formed on the ProteinChip array, researchers can use the ProteinChip array for two
unique purposes: 1) to identify proteins bound to the antibody on the basis of
molecular weight using the ProteinChip SELDI reader, or 2) to conduct epitope
discovery experiments by proteolytically cleaving proteins captured on ProteinChip
arrays (on-chip proteolysis) and then using SELDI to highlight the peptide(s)
associated with the epitope, streamlining the process of sequencing the protein
binding site via tandem mass spectrometry (MS/MS).
Fig. 1. Principles of the ProteinChip antibody capture kit. Protein G is covalently bound to the ProteinChip
PG20 array surface. An optional crosslinking step can be performed to crosslink IgG antibodies to protein G.