18 1 δ9 cis pc dopc

MEM media was purchased from Life Technologies (Grand Island, NY). G418 Disulfate was obtained from Caisson Labs (G030-5GM). Fura-2AM was purchased from Molecular Probes (Eugene, OR). DNP₂₅-BSA was from ThermoFisher Scientific (Waltham, MA; catalogue #A23018), custom DF3 peptide synthesized by and purchased from Anaspec (Fremont, CA) and anti-DNP-IgE was affinity-purified from ascites (Covance, Denver, PA) according to the methods of Liu et al (28 ). AlexaFluor647- (Thermofisher Scientific #A-20006), Janelia Fluor 646- (Tocris #6148) and CF640R- (Biotium #92108) labeled IgE lots were prepared using NHS Ester conjugation. Antibodies used are listed as follows: anti-pY PY20/99 cocktail (SantaCruz; sc-508, sc-7020), anti-rabbit-HRP secondary (SantaCruz; sc-2004, Cell Signaling #7074), anti-FcεRIγ (EMD Millipore; 06–727), Goat anti-rat IgE (Abnova; PAB29749). Poly-L lysine was purchased from Sigma-Aldrich (P1524-100MG), 18:1 (Δ9-Cis) PC (DOPC) and 16:0 DNP Cap PE lipids were purchased from Avanti Polar Lipids (850375C and 810528C, respectively). DNP-Peg12-NHS ester was purchased from BroadPharm (BP-22397). All live-cell RBL imaging was performed in modified Hank’s buffered salt solution (additional 10 mM Hepes, 0.05% w/v BSA, 5.45 mM glucose, 0.88 mM MgSO₄, 1.79 mM CaCl₂, 16.67 mM NaHCO₃).