Vectastain abc ap standard kit

Melanoma tissues were sampled from patients at the Pathology Department and Oncology Department, The Affiliated Hospital of Southwest Medical University in Luzhou, Sichuan, China. Expert pathologist (WQ) and dermatologist (CW) resected the primary tumors and the metastases and confirmed clear margins on the samples. For the analysis of PLA1A, desmocollin 3 (DSC3), and DMKN, the representative tissue sections from all three groups were transferred into 10% neutral buffered formalin; fixed and embedded in Tissue Tek II OCT (Miles Scientific, Naperville, IL, USA); frozen for 15 min in isopentane; pre-cooled in liquid nitrogen; and finally stored at −80 °C for future hematoxylin and eosin (H&E) staining and immunostaining assays. Hematoxylin (HHS16, Sigma-Aldrich) and eosin (HT110232, Sigma-Aldrich) staining of samples was performed according to the manufacturer's instructions. The best frozen sample was used for immunostaining of PLA1A, DSC3, and DMKN, which was performed following the streptavidin-biotin alkaline phosphatase complex method using the Vectastain ABC-AP standard kit (Vector Laboratories; Burlingame, CA) as previously described (59,60). Complete information about the primary monoclonal antibodies and secondary monoclonal antibodies has been provided in Supplementary Table S3.