Proteins were analyzed by SDS-PAGE using the NuPAGE system (ThermoFisher Scientific). Separated proteins were either identified by mass-spectrometric analysis or electrotransferred onto a nitrocellulose membrane by tank blotting with transfer buffer (ThermoFisher Scientific) according to the manufacturer’s instructions. The membranes were blocked with Universal Blot Buffer plus (Euroimmun) for 15 min and incubated with human serum or the polyclonal antibody against ITPR1 in Universal Blot Buffer plus for 3 h, followed by three washing steps with Universal Blot Buffer (Euroimmun), a second incubation for 30 min with anti-rabbit IgG-AP (Sigma-Aldrich), three washing steps, and staining with NBT/BCIP substrate (Euroimmun).
Universal blot buffer plus
Manufactured by EUROIMMUN
Universal Blot Buffer plus is a buffer solution designed for use in Western blot analysis. It is a concentrated formulation that can be diluted for the blocking, washing, and antibody incubation steps of the Western blot protocol.
Automatically generated - may contain errors
1 protocol using universal blot buffer plus
1
Western Blot Protein Analysis
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