Extended Data Fig. 1: SDS–PAGE analysis on SPEAR formation.
From: Antibody stabilization for thermally accelerated deep immunostaining
a–c, Establishment of P3PE-crosslinked IgG–Fab complex electrophoretic patterns and initial optimization of reaction conditions for yield. a, Reducing and b, non-reducing SDS-NuPAGE analysis of P3PE-crosslinked IgGs, Fabs and their complexes under various conditions and their electrophoretic patterns. c, Time course of P3PE crosslinking of IgG–Fab complexes. The tested reaction conditions are listed on the right. All experiments were repeated twice with similar results. d–g, Testing the tolerance of P3PE-crosslinking reaction towards common additives in commercially supplied antibodies using reducing SDS–PAGE. d, Screening for additives that inhibit P3PE crosslinking of IgG–Fab complexes. e, and f, Titration of Tris (e) and BSA (f) and their effects on P3PE-crosslinking reaction. The tested reaction conditions are listed on the right. g, Testing the effects of various fluorophores on the P3PE-crosslinking reaction. All experiments were repeated twice with similar results.
