F4 80 clone d2s9r

Manufactured by Cell Signaling Technology

F4/80 (clone D2S9R) is a monoclonal antibody that recognizes the F4/80 antigen, a transmembrane protein expressed on the surface of macrophages and monocytes. This antibody can be used for the identification and characterization of macrophages in various experimental systems.

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Lab products found in correlation

Ar6 antigen retrieval buffer, by PerkinElmer (1 mentions) Opal 4 color ihc kit, by PerkinElmer (1 mentions) Cd8 clone d4w2z, by Cell Signaling Technology (1 mentions) Dab substrate chromogen system, by Agilent Technologies (1 mentions) Citrate buffer ph 6, by Agilent Technologies (1 mentions)

2 protocols using f4 80 clone d2s9r

Profiling Tumor Immune Landscape

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Slides were stained using Opal 4-color IHC Kit (NEL794001KT) from Perkin Elmer along with primary antibodies from Cell Signaling Technology for CD3 (clone D4V8L), CD4 (clone D7D2Z), FoxP3 (clone D608R), CD8 (clone D4W2Z), CDllc (clone D1V9Y), F4/80 (clone D2S9R), and CD31 (clone D8V9E). Formalin-fixed paraffin embedded tissues were sectioned, deparaffinized in xylene, and rehydrated through an ethanol gradient. Microwave treatment was applied to perform antigen retrieval, quench endogenous peroxidases, and remove antibodies from earlier staining procedures. Perkin Elmer AR6 Antigen retrieval buffer (pH 6) was used for all antibodies. The slides were scanned with the VECTRA image scanning system (Perkin Elmer), and signals were unmixed into a composite image with Vectra inForm software. For each tumor model, 5 independent tumors were imaged with at least 10 images per slide from arbitrary fields of view. Images showing significant necrosis were not used for analysis. Final quantification was performed using custom MATLAB scripts.

McGrail D.J., Garnett J., Yin J., Dai H., Shih D.J., Lam T.N., Li Y., Sun C., Li Y., Schmandt R., Wu J.Y., Hu L., Liang Y., Peng G., Jonasch E., Menter D., Yates M.S., Kopetz S., Lu K., Broaddus R., Mills G.B., Sahni N, & Lin S.Y. (2020). Proteome instability is a therapeutic vulnerability in mismatch repair deficient cancer. Cancer cell, 37(3), 371-386.e12.

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Immunohistochemical Analysis of F4/80 Expression

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Immunohistochemistry was performed on formalin-fixed, paraffin-embedded tissue specimens. Briefly, 5 μm tissue sections were deparaffinized in Histoclear (Brunschwig, Basel, Switzerland) and rehydrated in descending concentrations of ethanol. Antigen retrieval was performed using citrate buffer, pH 6.0 (DAKO, Glostrup, Denmark) for 30 min at 98 °C. Endogenous peroxidases were blocked by incubation with 0.9% hydrogen peroxide for 15 min at room temperature (RT), blocking was performed using 3% bovine serum albumin for 1 h at RT in a wet chamber. Samples were stained for 1 h at RT with primary antibody F4/80 (clone D2S9R, Cell Signaling, #70076S), and for 1 h at RT with HRP-labeled secondary antibody. Antibody binding was visualized using a liquid DAB+ substrate chromogen system (DAKO). Then, samples were counterstained with hematoxylin and dehydrated in ascending ethanol solution and Histoclear.

Hering L., Katkeviciute E., Schwarzfischer M., Niechcial A., Riggs J.B., Wawrzyniak M., Atrott K., van de Sande M., Lang S., Becher B., Rogler G., Scharl M, & Spalinger M.R. (2021). Macrophages Compensate for Loss of Protein Tyrosine Phosphatase N2 in Dendritic Cells to Protect from Elevated Colitis. International Journal of Molecular Sciences, 22(13), 6820.

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