Light Regulation of Brassinosteroid Signaling Components: Checking Regulation of Protein Stability in Darkness.

Environmental conditions can affect stability of proteins at transcriptional or posttranscriptional levels to modulate their functions. Here we describe a method to observe changes in protein stability under different light conditions. In brief, Arabidopsis thaliana seedlings were maintained under various light regimes from continuous light to total darkness or transitions from light to dark, whereafter total protein was extracted from plants. Proteins were measured and resolved on sodium dodecyl sulfate-polyacrylamide gels and transferred to polyvinylidene difluoride membranes. Blots were incubated with the corresponding antibodies for the visualization of protein bands. The protocol described has been successfully applied in wild-type, different transgenic, and mutant background plants to study how light alone or in combination with other factors influences protein stability.