Extracted tumors were fixed with 10% formalin, embedded in paraffin and sliced in 4 μm thickness. The multiplex IHC was performed with an Opal 7-Color Automation IHC Kit (Akoya Biosciences, Hopkinton, MA, USA) and BOND RXm automated stainer (Leica Biosystems, Wetzlar, Germany) using the following antibodies: anti-pan-cytokeratin (CK) (rabbit poly; Bioss, Woburn, MA, USA), anti-CD8α (EPR20305; Abcam, Cambridge, United Kingdom), anti-CD44 (IM7; Bio X Cell, Lebanon, NH, USA), anti-F4/80 (D2S9R; Cell Signaling Technology, Danvers, MA, USA), anti-Ly6G (1A8; Bio X Cell, Lebanon, NH, USA), anti-CD11b (EPR1344; Abcam, Cambridge, United Kingdom) and anti-Gzmb (rabbit polyclonal; Abcam, Cambridge, United Kingdom). The slides were imaged in the Mantra Quantitative Pathology Workstation (Akoya Biosciences, Menlo Park, CA, USA). The multiplex IHC images were analyzed using inForm Tissue Finder software (Akoya Biosciences, Menlo Park, CA, USA). Cell phenotype was defined based on the antigen expressions as the following: CD8+ = CD8+ T cell, F4/80+ = macrophage, Ly6G+CD11b+ = neutrophil, Gzmb+CD3− = NK cell. Tissue phenotype was determined as the following: pan-cytokeratin positive = tumor. CD8+ T cells in the tumor area were counted as tumor-infiltrating CD8+ T cells.
Anti f4 80 d2s9r
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-F4/80 (D2S9R) is a laboratory research use antibody that recognizes the F4/80 antigen. The F4/80 antigen is a cell surface glycoprotein expressed on mature mouse macrophages.
Automatically generated - may contain errors
Lab products found in correlation
Anti ly6g 1a8, by BioXCell (1 mentions)
Opal 7 color automation ihc kit, by Akoya Biosciences (1 mentions)
Mantra quantitative pathology workstation, by Akoya Biosciences (1 mentions)
Inform tissue finder software, by Akoya Biosciences (1 mentions)
Anti pan cytokeratin ck rabbit poly, by Bioss Antibodies (1 mentions)
Anti cd8α epr20305, by Abcam (1 mentions)
Anti cd11b, by Abcam (1 mentions)
Mach2 double stain 1, by Biocare Medical (1 mentions)
Aperio at2, by Leica (1 mentions)
Vectastain abc kit, by Vector Laboratories (1 mentions)
2 protocols using anti f4 80 d2s9r
1
Multiplex IHC for Tumor Immune Profiling
2
Immunohistochemical Analysis of Xenograft Samples
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Serial 4-μm-thick paraffin sections from formalin-fixed, paraffin-embedded xenografts, obtained as described above (see In vivo studies) and as previously reported (8, 11 (link)), were processed according to standardized IHC procedures and then immunostained with the following antibodies: anti-CD47 (clone EPR21794; #ab218810, rabbit monoclonal, Abcam), anti-F4/80, (D2S9R) (#70076, rabbit polyclonal, Cell Signaling, RRID: AB_2799771), anti-CD206 (#ab64693, rabbit polyclonal, Abcam, RRID: AB_1523910), and anti-CD99 (O13) (#915601; BioLegend, RRID:AB_2565169). An avidin–biotin–HRP method was used for single staining (VECTASTAIN ABC kit, PK-4001, Vector Laboratories). A polymer with anti-mouse alkaline phosphatase and anti-rabbit HRP was used for dual staining (Mach2 Double Stain #1, #PBC-MRCT523L, Biocare Medical). For morphologic analyses, the slides were stained with hematoxylin and eosin.
Histologic and histomorphometric analyses were carried out with a digital pathology slide scanner with a resolution of 0.5 μm/pixel (Aperio AT2, Aperio Technologies, Vista, CA) using ImageScope software (v12.4.3, RRID: SCR_014311) for slide viewing and analysis.
Histologic and histomorphometric analyses were carried out with a digital pathology slide scanner with a resolution of 0.5 μm/pixel (Aperio AT2, Aperio Technologies, Vista, CA) using ImageScope software (v12.4.3, RRID: SCR_014311) for slide viewing and analysis.
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