Antibody Validation as a tool to study “Synaptic Adhesion Proteins”

Abstract

Historically there has been an absence of reliable and validated antibodies against synaptic adhesion proteins, particularly neurexins and neuroligins. This has led to the failure of the vital experiments that study the function of these synaptic proteins, leading to wastage of time and expensive resources. My project validated antibodies against neurexins and neuroligins through the process of immunoprecipitation, which pulls the protein of interest, followed by the process of western blot to visualize the results. The results indicated that the antibodies against β isoforms across the three neurexins performed better. Similarly, antibodies against Nlgn 2 and Nlgn 3 showed consistent bands congruent with the molecular weight of neuroligin. This project also investigated the role of Nrxn1γ, an isoform of neurexin1, in maintaining the excitatory-inhibitory balance between synapses that is required for optimal synaptic function. Collectively, these findings not only establish a dependable set of antibodies for probing neurexin and neuroligin biology but also underscore Nrxn1γ as a meaningful contributor to inhibitory synapse maintenance, strengthening both the technical and conceptual foundations for future synaptic research.