Anti pd 1 mouse antibody 315m

Recently, TILs, PD-1, and PD-L1 were investigated in this STS cohort [9 (link)]. We used those results to explore the relationship between VISTA and the PD-1/PD-L1 pathway in STS. TILs between tumour cells were counted per high-power field (HPF) (400× magnification, field of view 0.237 mm²) in H&E-stained TMA slides, as routinely carried out by the pathologist.
As described previously [9 (link)], slides were pre-treated with heat and Target Retrieval solution (S1699, Agilent, Santa Clara, CA, USA) before incubation with the monoclonal primary anti-PD-1 mouse antibody (315M; 1:80; Cell Marque, Rocklin, CA, USA) for 60 min at room temperature. The Vectastain Elite ABC HRP Kit (Vector Laboratories, Burlingame, CA, USA) and the chromogen DAB+ (Agilent) were used for detection and Hematoxylin (Vector Laboratories) for counterstaining.
For PD-L1 staining, slides were pre-treated with heat and the Epitope Retrieval Solution pH8 Novocastra (Leica Biosystems, Wetzlar, Germany) before incubation with the monoclonal primary anti-PD-L1 rabbit antibody (E1L3N; 1:50; Cell Signalling Technology) for 60 min at room temperature.
For CD3 staining, a monoclonal antibody raised in rabbit (SP7; 1:150; Zytomed, Berlin, Germany) was employed according to standard procedures. We used the SignalStain Boost IHC Detection Reagent (Cell Signalling Technology) and the chromogen DAB+ (Agilent) for detection.

Tumor-infiltrating lymphocytes (TILs), PD-1 and PD-L1 were previously investigated in our HR-STS cohort [38 (link),39 (link)]. TILs were counted per high-power field (HPF) (400× magnification, field of view 0.237 mm²) in H&E-stained TMA slides. As previously described, slides were pre-treated with heat and Target Retrieval solution (S1699, Agilent, Santa Clara, CA, USA) before incubation with the monoclonal primary anti-PD-1 mouse antibody (315M; 1:80; Cell Marque, Rocklin, CA, USA) for 60 min at room temperature. The Vectastain Elite ABC HRP Kit (Vector Laboratories, Burlingame, CA, USA) and the chromogen DAB+ (Agilent) were used for detection, and Hematoxylin (Vector Laboratories) for counterstaining. For PD-L1 staining, slides were pre-treated with heat and the Epitope Retrieval Solution pH8 Novocastra (Leica Biosystems, Wetzlar, Germany) before incubation with the monoclonal primary anti-PD-L1 rabbit antibody (E1L3N; 1:50; Cell Signaling Technology, Danvers, MA, USA) for 60 min at room temperature. We used the SignalStain Boost IHC Detection Reagent (Cell Signalling Technology) and the chromogen DAB+ (Agilent) for detection according to previous studies [39 (link)].