Multiplex IHC was performed as previously described (34 (link)), using Opal Automation IHC Kit (Akoya Bioscience) and Bond RXm autostainer (Leica Biosystems). The following antibodies were used: anti-pan cytokeratin (rabbit poly; RRID: AB_10855057, Bioss), anti-PDPN (clone 66; RRID: AB_2785565, Invitrogen), anti-αSMA (clone EPR5368; RRID: AB_11129103, Abcam), anti-CD31 (clone D8V9E; RRID: AB_2722705, Cell Signaling Technology), anti-VEGFR3 (clone AFL4; RRID: AB_467795, Thermo Fisher Scientific), and anti-digoxigenin (DIG, clone 9H27L19; RRID: AB_2532342, Thermo Fisher Scientific; 1:500). Cells were counterstained with DAPI. Stained slides were imaged with Mantra Quantitative Pathology Workstation (Akoya Biosystems), and images were analyzed with inForm software (RRID: SCR_019155, Akoya Biosystems).
Anti pan cytokeratin rabbit poly
Manufactured by Bioss Antibodies
Sourced in United States
Anti-pan cytokeratin (rabbit poly) is a laboratory reagent used to detect the presence of cytokeratins, a group of proteins found in the cytoplasm of epithelial cells. This polyclonal antibody, derived from rabbit, is designed to recognize multiple cytokeratin subtypes, providing a broad-spectrum detection of these structural proteins.
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2 protocols using anti pan cytokeratin rabbit poly
1
Multiplex IHC for Tissue Characterization
2
Multicolor Immunofluorescence Analysis of TILs
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Multicolor immunofluorescence staining was performed to analyze the tumor-infiltrating lymphocytes (TILs) as previously described [12 (link),13 (link)]. The sections were stained with DAPI and the following antibodies: anti-CD8 (clone EPR20305; Abcam, Cambridge, UK; cat # ab209775; 1:500 dilution), anti-CD4 (clone EPR 19514; Abcam; cat # ab221775; 1:1000 dilution), anti-Foxp3 (clone 1054C; Novus Biologicals, Centennial, CO, USA; cat # MAB8214; 1:1000 dilution), and anti-pan cytokeratin (rabbit poly; Bioss Antibodies, Woburn, MA, USA; cat # bs-1712R; 1:250 dilution). Tissue area was divided into “Tumor” and “Stroma” based on the expression of pan cytokeratin. Four pictures were obtained for each specimen, and tissue area and cell count were summed for each tissue category. Cell density was calculated as cell counts per square millimeter.
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