10/08/2026
This summer, Caden researched the importance of lysis buffer choice in Dr. Aldrin V. Gomes' lab.
"Western blotting, also known as immunoblotting, is a process in which specific proteins are able to be detected and quantified. Despite having existed for several decades, further refinement in order to increase repeatability and clarity is still needed, as variability in sample preparation remains an important source of experimental inconsistency. My work concentrated on how different formulations of radioimmunoprecipitation assay buffers (RIPA buffers) affect the final western blot signal and protein extraction. Five commonly used commercial RIPA buffer formulations were compared using liver tissue from 5-month-old female mice (n=3). Following tissue homogenization, protein quantification, SDS-PAGE, electrotransfer to nitrocellulose membranes, and enhanced chemiluminescent detection, protein bands for PSMA1 and Rpt6 were quantified with ImageJ and normalized to total protein via Ponceau S staining. One-way ANOVA showed a significant effect of buffer formulation on Rpt6 signal, but not on PSMA1 signal, suggesting that the influence of buffer composition may depend on the target protein. Overall, findings demonstrate that RIPA buffer composition can measurably influence immunoblot signal."