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After three 15-minute washes with phosphate-buffered saline with tween-20 (PBST), the membranes were incubated with horseradish peroxidase-conjugated goat anti-rabbit IgG secondary antibody (1:2000 dilution, ab205718, Abcam, Shanghai, China) for 2 hours at room temperature

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Diabetologia 62:10361047 Pugliese A, Yang M, Kusmarteva I, Heiple T, Vendrame F, Wasserfall C, Rowe P, Moraski JM, Ball S, Jebson L, Schatz DA, Gianani R, Burke GW, Nierras C, Staeva T, Kaddis JS, Campbell-Thompson M, Atkinson MA (2014) The Juvenile Diabetes Research Foundation Network for Pancreatic Organ Donors with Diabetes (nPOD) program: goals, operational model and emerging findings

If you develop severe allergic reactions such as difficulty breathing, facial swelling, or widespread rash after a B12 injection, call 999 or go to A&E immediately, as these may indicate anaphylaxis, though this is extremely rare