Intercept tbs antibody diluent buffer

Western blot analysis was performed to determine the levels of S glycoprotein for each of the VSV pseudoviruses. 30 μL of virus is obtained and mixed with 10 μL of 4x LDS sample reducing buffer. Samples were boiled at 95 °C for 5 minutes and spun down. 35 μL of final mixture was run on 4–20% Mini-Protean TGX precast protein gel at 150 Volts for 45 minutes. Proteins were transferred using Trans-Blot Turbo Mini 0.2 μm PVDF Transfer Packs and membranes were incubated with Intercept (TBS) Blocking Buffer (Licor) for 1 hour at room temperature. Anti-Fusion peptide antibody at 1 μg/ml was diluted in Intercept (TBS) Antibody Diluent Buffer (Licor) and incubated at 4°C overnight. The following day membranes were washed with TBST 3x with 5-minute incubations at room temperature. Anti-human Fc (Jackson ImmunoResearch) was diluted 1:50,000 in Intercept (TBS) Antibody Diluent Buffer (Licor) and membranes were incubated at room temperature for 1 hour. Membranes were washed with 3x TBST with 5 minute incubations at room temperature. Images were taken using the Licor Odyssey CLx.