From the crude polyclonal the crossreactive antibodies were extracted by incubation with Sepharose-bound human IgM and IgG.
Specific antibodies were absorpted by incubation with Sepharose-bound human IgA.
Specific antibodies were eluted by acidic buffer at pH 2.5 followed by neutralisation and dialysis.
After repeated binding with immobilized human IgA a minimum of 65% protein bound.
The purified polyclonal was conjugated to horseradish peroxidase according to the periodate method followed by gel-filtration and ion-exchange chromatography to clear unbound conjugate.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
This product has been referenced in:
Kempsell KE et al. Whole genome protein microarrays for serum profiling of immunodominant antigens of Bacillus anthracis. Front Microbiol6:747 (2015).
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Csorba K et al. Development of an ELISA for sensitive and specific detection of IgA autoantibodies against BP180 in pemphigoid diseases. Orphanet J Rare Dis6:31 (2011).
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