Application of the dot immunobinding assay to allergy diagnosis

Immunoblotting and dot blotting

“…Immunoblotting is ideally suited to this kind of problem and has considerable potential for identification, purification and investigation of the structure and properties of allergens. Identification of IgE antibodies against defined allergens by dot blotting (Derer et al, 1984;Singh and Knox, 1985) could well prove to be a useful technique in the clinical immunology laboratory for diagnosis of hypersensitivity reactions. Several Sutton et al (1982); Ford et al, (1985Ford et al, ( , 1986; Mecheri et al (1985); Singh and Know (1985); Haas et al (1986); Alterman et al (1987) lpsen and Larsen (1988) Einarsson (1987); Hoffman (1987) Kroutil and Bush (1987) immunoblotting studies have been published (Table IV) but, in view of the enormous variety and complexity of allergens, there is clearly much more to be done.…”

Monoclonal anti‐IgE antibodies in the diagnosis of dog allergy

“…In all experiments a dot‐immunobinding technique was used (unless otherwise mentioned). This method has been described in detail elsewhere 4 . 5…”

Oral anti-IgE immunization with epitope-displaying phage

“…To determine the titer of remaining infective phage particles, fresh E. coli XL-1 blue were infected for 15 min, and aliquots titrated on agar plates containing appropriate antibiotics. The effect of the acid treatment on the immunologic reactivity of the immunizing Ag was analyzed in an immunodot assay [35]. Nitrocellulose strips (Millipore Corp., Bedford, MA) were coated with 1 ?…”

“…Aliquots of acid-treated phages were incubated overnight and detected with polyclonal HRPlabeled anti-phage antibody (1 : 1000 in TBS-C) for 4 h. Strips were developed as described [32]. The OD of the dots was measured using a hand-held Gretag densitometer (Gretag Limited, Regensdorf, Switzerland) and are expressed as relative OD (rel OD) (see [32,35]). …”