Analyzing Protein Changes in Guinea Pig Tissue Lysates Using Non-guinea Pig Specific Antibodies: Procedures for Western Blotting and Examples Using 16 Individual Antibodies for Common CNS Proteins

摘要

Behavioral studies investigating chemical weapon nerve agent (CWNA) exposure often utilize guinea pigs because their levels of carboxylesterase enzymes are similar to humans (Maxwell et al. 1987; Shih and McDonough 1999; Shih and McDonough 2000; de Groot et al. 2001). However, the guinea pig model does present a significant problem when trying to correlate behavioral and protein changes due to the absence of guinea pig-specific antibodies. We have developed a procedure to determine the specificity of commercially available, non-guinea pig-specific antibodies in guinea pig lysates. Common Western blotting techniques were used to compare immunostaining patterns of tissue lysates between a known species, rat, and the guinea pig using antibodies to several common CNS proteins. Of the sixteen antibodies tested, nine revealed the exact same banding patterns as their rat lysate controls, five had similar banding patterns but required further characterization and two revealed no immunoreactivity. This report outlines the procedure for characterizing the immunoreactivity of these antibodies for use in guinea pig tissue lysates and identifies nine CNS proteins that can be labeled in this manner. This procedure should be of benefit to all investigators using the guinea pig behavioral model who wish also to investigate any underlying protein changes.