Generation and characterization of murine monoclonal antibodies specific for XXXX immunoglobulin E.

@entity291 @entity214 @entity2975
Monoclonal antibodies were produced against serum-derived @entity291 immunoglobulin E (IgE). Culture supernatants of hybridomas were initially screened by enzyme-linked immunosorbent assay (ELISA). Supernatant-derived antibodies were concentrated and further characterized using ELISA, reverse cutaneous anaphylaxis, immunohistochemical staining, and immunoblotting of IgE-containing samples separated by @entity214 - @entity2975 gel electrophoresis (PAGE). Eight monoclonal antibodies showed specificity for @entity291 epsilon immunoglobulin heavy chain. Two antibodies (E2 and E32) reacted in immunoblots of @entity214 -PAGE of serum IgE under reducing conditions. Additionally, E2, E5, and E32 detected epsilon chain in serum separated by @entity214 -PAGE and then renatured. Antigen-specific IgE was detected in Western blots by E5 and E32. Immunoperoxidase staining of IgE-containing cells in mesenteric lymph node sections was detected with E5, E21 and E32. All eight antibodies produced positive reverse cutaneous anaphylaxis reactions in @entity291 skin. All functioned well in ELISA as a plate-sensitizing reagent for quantitation of total IgE; E5 and E32 worked well as a primary antibody in antigen-specific IgE assays. These antibodies will be useful in research applications and in diagnostic assays.
