Troubleshooting using anti-light chain antibodies
Anti-light chain antibodies are commonly used for Western blotting (WB) after Immunoprecipitation (IP) when detecting proteins that migrate to approximately 50kDa with IP and WB primary antibodies from the same host species. These proteins would otherwise be obscured by detection of the heavy chain from the precipitating antibody. Unlike whole molecule-specific antibodies, which detect both heavy and light chains, the anti-IgG, Light Chain Specific antibodies only detect light chains. Consequently, the native primary antibodies used for WB are detected by recognition of their light chains, while the heavy chain of the IP antibody is not detected. Consequently, proteins of interest of approximately 50 kDa can be resolved on a blot without an overlapping signal from the IP antibody heavy chain.