Anti cgas clone d1d3g

Proximity ligation assay (PLA) was performed using NaveniBright HRP kit (Navinci Diagnostics) according to the manufacturer’s protocol using the following primary antibodies: anti-cGAS (clone D1D3G, #15102, 1:100 pH9, Cell Signaling) and anti-DNA double stranded (clone AE-2, MAB1293, 1:100 pH9, Abcam). Negative controls were performed using only one primary antibody. Slides were analyzed under a Zeiss Axioscope A1 and microphotographs were collected using a Zeiss Axiocam 503 Color with the Zen 2.0 Software (Zeiss). Sections were subsequently immunostained to detect the expression of the E2.2 antigen. IHC was developed using SignalStainBoost IHC Detection rabbit (cod. #18653, Cell Signaling Technology) alkaline phosphatase-conjugated produced in horse and Vulcan Fast Red as substrate chromogen. Again, the sections were analyzed and photographed using Zeiss Axioscope A1 and Zeiss Axiocam 503 Color with the Zen 2.0 Software (Zeiss). Segmented images were obtained using HALO image analysis software (v3.2.1851.229, Indica Labs). HALO image analysis software was used to quantify the PLA signals in twelve non-overlapping fields at high-power magnification (400X) and the output was expressed as “percentage positive cells”.