The Thermo Scientific Pierce A IgG Plus Orientation Kit uses a crosslinker to covalently attach antibody that is bound to Protein A agarose resin, thereby creating a reusable affinity column for purification of antigens.
Features of the Protein A IgG Plus Orientation Kit:
• Contaminant-free antigen purification—provides for antigen purification without contamination (co-elution) of antibody • Covalent, leak-resistant antibody immobilization—high-capacity binding of IgG to Protein A agarose, followed by efficient crosslinking with DSS crosslinker • High-quality agarose resin—crosslinked 6% beaded agarose with recombinant Protein A • Gentle binding and crosslinking conditions—coupling antibody in standard PBS • Oriented immobilization—binds and crosslinks IgG with the antigen-binding sites directed outward, able to capture passing antigen in the mobile phase without steric hindrance • Complete kit—includes all the reagents needed to immobilize the antibody • Better than DMP—less antibody leaching than traditional orientation method with DMP crosslinker
This 'Plus' kit provides for more efficient and stable antibody immobilization than the traditional orientation method because it uses the NHS-ester crosslinker DSS (disuccinimidyl suberate) rather than the imidoester crosslinker DMP (dimethyl pimelimidate). Several milligrams of IgG can be bound and crosslinked to each 2 ml column, after which the prepared affinity column can be used multiple times for antigen purification using typical binding and elution conditions.