Student Scholars Day

Purification of cAR1 Antibody in Dictyostelium discoideum

Henry Hall Atrium 97

Dictyostelium discoideum is an organism that exhibits cell signaling, movement, and development. When starved, these cells secrete cyclic adenosine phosphate (cAMP) to signal nearby cells, increasing expression of the cAMP receptor protein (cAR1). In past BIO 406 classes, an anti-cAR1 antibody was used to detect the receptor protein in these cells; however, western blots showed high background. Our goal was to purify this antibody batch to reduce the background. After initiating expression of cAR1, proteins were extracted and separated on a SDS-PAGE gel. Proteins were visualized on a western blot using the cAR1 antibody (predicted size 44kD). We found that three uses of the antibody was the optimal number of absorptions to reduce non-specific background. However, there was only one band present in both starved and unstarved cells at 120kD, indicating that it was not the cAR1 protein. Ultimately, we concluded that this antibody batch would not be useful for future BIO 406 experiments.