08 February 2021 4 4K Report

Hello,

I am western blotting SARS-CoV-2 spike protein from lentiviral using an antibody against S1. To prepare samples, I mixed concentrated lentivirus 1:1 with 2X Laemmli buffer and boiled for 10 minutes.

In my western, I see a large band 130kDa which corresponds with S1+S2 as well as a band at 100kDa which corresponds with S1. I also see two large bands between 50 and 60kDa. What could these bands correspond to? I don't think that the antibody should recognize S2. Are these possibly products of S1 denaturation from my boiling and buffer? Would you recommend running the protein in non-reducing conditions?

Thank you.

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