Fkbp51 goat polyclonal antibody

A Duolink In Situ PLA Detection Kit (Sigma-Aldrich) was used to detect the interaction between FKBP51 and PR. 4% PFA-fixed paraffin-embedded decidua basalis specimens from women with iPTB (n = 6) versus GA-matched control PA or PE (n = 6/each) were deparaffinized and rehydrated, and then, antigen retrieval was carried out by incubating the slides in citrate buffer (pH = 6.0) in microwave for 20 min. After washing steps, the slides were incubated with blocking solution for 1 h at 37 °C and then incubated with primary antibodies (PR rabbit monoclonal antibody; Cell Signaling and FKBP51 goat polyclonal antibody; R&D Systems) overnight at 4 °C. Primary antibodies were carefully selected from different species sources based on negligible background. Hybridization with anti-rabbit MINUS and goat PLUS PLA probes, ligation, and amplification of conjugants were performed as per the manufacturer’s instructions. Negative controls were included in all experiments in the absence of primary antibodies. Slides were mounted with in situ mounting medium with DAPI (Sigma-Aldrich). A total of 10 different areas were randomly selected per slide, and photographs were obtained with 40× or 100× magnification, and images were analyzed using a ZEISS fluorescence microscope with the ZEN 2012 software system. The interaction of FKBP51 with PR was calculated as the number of PLA-positive dots per cell.