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. 2021 May 5;76(10):3028–3040. doi: 10.1111/all.14832

FIGURE 1.

FIGURE 1

Detection of allergen sensitization by quantifying binding of fluorescent allergen components of BV and RGP to basophils. A) Schematic diagram of DNA constructs for generation of the recombinant allergens Api m 1, Lol p 1, and Lol p 5. Recombinant allergens contained mutations to render the catalytic site inactive (Api m 1 H67Q, Lol p 1 H104V). Leader peptide and spacer (L); 6‐histidine tag (His); avidin tag (AviTag). B) Western blots of recombinant allergens detected with anti‐His antibody. C) Representative flow plot for detection of circulating basophils (CD123+IgE+) within gated mononuclear cells (Figure S1). D) Histograms depicting representative fluorescent staining of Api m 1, Lol p 1, and Lol p 5 tetramers on basophils from relevant allergic patients (red) and controls (blue). Values above the histograms represent median fluorescence intensity values of the population.