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Tordesillas, Leticia, PhD; Rahman, Adeeb H., PhD; Hartmann, Boris M., PhD; Sampson, Hugh A., MD; Berin, M. Cecilia, PhD
Journal of allergy and clinical immunology, 12/2016, Volume: 138, Issue: 6Journal Article
Whole blood from peanut-allergic patients (n = 6) or healthy controls (n = 3) was stimulated with peanut extract, anti-IgE, or media control for 15 or 30 minutes before fixation and red blood cell lysis. There was also increased phosphorylation of S6 (Fig 1, G), which was also observed in monocytes, cDCs, and pDCs (Fig 2, A and B) of peanut-allergic subjects but not healthy controls in response to peanut, and in all subjects in response to anti-IgE.\n Then, samples were first washed and permeabilized with ice-cold methanol for 30 minutes and then washed and stained with intracellular phosphoprotein antibodies for 30 minutes on ice, with prior heparin blocking.
