Mab102.1f10 and control igg4

EF‐hand allergens (Phl p 7, Aln g 4, Bet v 4, Bra r 1, Che a 3, Ole e 3, Ole e 8) and BSA (negative control) were dotted to nitrocellulose strips (1 μg/dot) (Schleicher & Schuell, Dassel, Germany) and incubated with sera from 14 Phl p 7‐allergic patients (diluted 1 : 5), with serum from a nonallergic individual, with mAb102.1F10 and control IgG₄ (1 μg/strip; Sigma‐Aldrich, St. Louis, MO, USA) and with Phl p 7‐specific rabbit antiserum and control antiserum raised against Der p 2 from house dust mite. Bound human IgG₄ antibodies were detected with a mouse monoclonal anti‐human IgG₄ antibody (PharMingen, San Diego, CA, USA), followed by a ¹²⁵I‐labelled rabbit anti‐mouse antiserum (Perkin Elmer, Waltham, MA, USA), whereas bound rabbit antibodies were detected with ¹²⁵I‐labelled goat anti‐rabbit antisera (Perkin Elmer). Bound patients' IgE antibodies were detected with ¹²⁵I‐labelled anti‐human IgE (BSM Diagnostica, Vienna, Austria) (data not shown). Bound ¹²⁵I‐labelled antibodies were quantified with a gamma counter (Wizzard, Automatic Gamma Counter; Wallac, Uppsala, Sweden), and results represent mean counts per minute (cpm)/dot (Fig. 1A).