Mouse Anti-Furazolidone Recombinant Antibody (clone SPE7) (CAT#: PABZ-048)

Figure 1 Inhibition of SPE7 Fv binding to immobilized DNP-Asn by various cross-reactants as measured by competitive ELISA.

Noted are the abbreviated names of the various cross-reactants, the full names and structures of which are given in Table 2. Residual SPE7 binding to immobilized DNP-Asn was determined by measuring capture of SPE7 using anti-mycHRP secondary antibody (see Materials and Methods). Binding, as a percentage of the binding observed in the absence of any inhibitor, was plotted against inhibitor concentration. IC₅₀s were determined as the concentration of ligand needed for 50% inhibition and are given in Table 2.

James, L. C., & Tawfik, D. S. (2003). The specificity of cross‐reactivity: promiscuous antibody binding involves specific hydrogen bonds rather than nonspecific hydrophobic stickiness. Protein Science, 12(10), 2183-2193.

Figure 2 SPE-7 IgE Fab inhibits SPE-7 IgE induced LAD-2 cell activation.

(A) Percentage degranulation of LAD-2 cells, induced by 30 nM SPE-7 IgE, was dose-dependently inhibited by addition of indicated molar excess of recombinant SPE-7 IgE Fab. (B) Data are shown in an inhibition curve. IC₅₀ of SPE-7 IgE Fab inhibition of SPE-7 IgE induced degranulation = 190 nM. Lower and upper dashed lines indicated cell-only background control and activation by SPE-7 IgE only, respectively. (C) LAD-2 cell TNF-α release, evoked by 30 nM SPE-7 IgE, was dose-dependently inhibited by addition of indicated molar excess of recombinant SPE-7 IgE Fab. (D) IC₅₀ of recombinant SPE-7 IgE Fab inhibition of SPE-7 IgE evoked TNF-α release ~170 nM. Dashed line indicates cell-only background control. Means of 4 to 10 independent experiments ± SEM are shown. Statistically significant difference to SPE-7 IgE only was determined by one-way ANOVA with Dunnett's post-test; *** p < 0.001, * p = 0.01 to 0.05, ns = not significant p ≥ 0.05.

Bax, H. J., Bowen, H., Dodev, T. S., Sutton, B. J., & Gould, H. J. (2015). Mechanism of the antigen-independent cytokinergic SPE-7 IgE activation of human mast cells in vitro. Scientific reports, 5, 9538.