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The plasmid was mixed 1:3 (w/w) with PEI MAX 40,000 (Polysciences, 24765-1) for 20Lmin at room temperature and then used to transfect Expi293 cells at 37C in 8% CO2 (Thermo A14527)

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Products are provided as lyophilized powder in sealed vials with documented peptide content (e.g., 5 mg or 10 mg)

After incubating with 5% bovine serum albumin for 1 h, the membranes were incubated with primary antibodies overnight at 4 C

Conflating the two in a study design would be a meaningful error a point covered further in our related article on common laboratory research mistakes