Generation and characterization of a rat monoclonal antibody specific for PCNA.

摘要

Proliferating cell nuclear antigen (PCNA) is a homotrimeric ring-shaped protein that encircles the DNA and acts as a stationary loading platform for multiple, transiently interacting partners participating in various DNA transactions. This essential cellular component, originally characterized as a nuclear antigen of dividing cells, is evolutionary highly conserved from yeast to human. Within the eukaryotic cell, PCNA serves as a processivity factor for DNA polymerase delta and plays a key role in DNA replication, repair, cell cycle regulation, and post-replicative transactions like DNA methylation and chromatin remodelling. All these cellular processes are regulated by a complex network comprising cell cycle dependent changes in expression levels, dynamics, interactions, and localization of PCNA. Here we report the generation and characterization of the first rat monoclonal antibody (MAb) against human PCNA, designated as PCNA 16D10. We demonstrated that PCNA 16D10 MAb has high affinity and specificity and is suited for ELISA, immunoblotting, immunoprecipitation, and immunofluorescence. The characteristic punctate staining of S phase cells allows the identification of proliferating cells and the monitoring of cell cycle progression.