F4 80 pe cy7

Manufactured by Thermo Fisher Scientific

Sourced in United States

The F4/80-PE-Cy7 is a fluorochrome-conjugated antibody used for the identification and quantification of the F4/80 antigen expressed on the surface of macrophages and monocytes. It can be used in flow cytometry applications to analyze the presence and distribution of these cell types in research samples.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

F4/80 (353 citations) F4/80-PE (78 citations) F4/80-PE-Cy7 (BM8, (4 citations) PE/Cy7-conjugated anti-mouse F4/80 (2 citations)

23 protocols using f4 80 pe cy7

Multicolor Flow Cytometry of Tumor Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Tumor cells were resuspended at a concentration of 10⁶ cells/100 µL and cell surface antigens were stained with fluorophore-conjugated antibodies: NIR-L/D (Invitrogen, Waltham, MA, USA, cat. 17-5321-81), BV510-CD45 (BD, Franklin Lakes, NJ, USA, cat. 563891), PE-Cy7-F4/80 (Invitrogen, cat. 25-4801-82), FITC-CD206 (BioLegend, San Diego, CA, USA, cat. 141704), APC-CD3 (eBioscience, San Diego, CA, USA, cat. 17-0031-82), FITC-CD4 (Thermo Fisher, Waltham, MA, USA, cat. 11-0041-82), PE-Cy5.5-CD8a (ThermoFisher, cat. 45-0081-42), and PE-CD25 (Invitrogen, cat. 12-0251-83). Antibody staining occurred for 30 min at 4 °C in the dark. Cells were acquired on the Attune NxT flow cytometer (Thermo Fisher). Compensation and sequential gating (see Supplementary Materials) were performed with FlowJo software (FlowJo LLC, v.10.8.1; Ashland, OR, USA).

Napier T.S., Lynch S.E., Lu Y., Song P.N., Burns A.C, & Sorace A.G. (2023). Molecular Imaging of Oxygenation Changes during Immunotherapy in Combination with Paclitaxel in Triple Negative Breast Cancer. Biomedicines, 11(1), 125.

+ Open protocol

+ Expand

Immunophenotyping Mouse Blood Cell Populations

Check if the same lab product or an alternative is used in the 5 most similar protocols

Blood was collected from mice via the retro-orbital sinus, and red blood cells were lysed before staining with the following fluorochrome-conjugated antibodies: BV650 CD45.1 (BioLegend clone A20), AlexaFluor700 CD45.2 (BioLegend clone 104), BUV496 B220 (BD Biosciences clone RA3-6B2), PerCP-Cy5.5 CD3e (BioLegend clone 145-2C11), APC-Cy7 CD11b (BioLegend clone M1/70), APC Ly6g (BioLegend clone 1A8), BV605 Ly6c (BioLegend clone HK1.4), BV421 Ter-119 (BioLegend clone TER-119), and PE-Cy7 F4/80 (Invitrogen BM8). Data were collected using a LSRII (BD Biosciences) and analyzed using FlowJo V10 (BD Biosciences). The following cell surface markers were used to isolate or phenotype the following cell types: hematopoietic stem cells (HSCs), Lin-Sca-1+ c-Kit+ Flt3-CD150+ CD48-; HSPCs, Lin-Sca-1+ c-Kit+; granulocyte/macrophage-primed multipotent progenitors (MPP G/M ), Lin-Sca-1+ c-Kit+ Flt3-CD150-CD48+; common myeloid progenitor cells (CMPs) Lin-Sca-1-c-Kit+ CD34+ FcγR-; and granulocyte-macrophage progenitors (GMPs), Lin-Sca-1-c-Kit+ CD34+ FcγR+. Data were collected using a BD FACSymphony A5 or cells were prospectively isolated using a FACSymphony S6 (BD Biosciences). All flow cytometry data were analyzed using FlowJo V10.

Schwartz L.S., Young K.A., Stearns T.M., Boyer N., Mujica K.D, & Trowbridge J.J. (2024). Transcriptional and functional consequences of Oncostatin M signaling on young Dnmt3a-mutant hematopoietic stem cells. Experimental hematology, 130.

+ Open protocol

+ Expand

Tumor Macrophage Profiling via Flow Cytometry

Check if the same lab product or an alternative is used in the 5 most similar protocols

To prepare single-cell suspensions for flow cytometry, fresh tumor tissue was dissected into approximately 1 to 3 mm³ fragments and digested with 80 U/mL collagenase (Invitrogen) in DMEM containing 10% FBS for one hour at 37°C while shaking. After red blood cell (RBC) lysis, single-cell suspensions were filtered and incubated for 20 minutes on ice with the following antibodies (1:100): CD45-PE (eBioscience, San Diego, CA), F4/80-PE-Cy7 (eBioscience, San Diego, CA), CD11b-FITC (BD Biosciences San Jose, CA), CD206-APC (Biolegend, San Diego, CA),CD68-PerCP-Cy5.5 for macrophages (Biolegend, San Diego, CA), Cells were washed with phosphate-buffered saline (PBS) before analysis on the BD LSR-II flow cytometer (Beckman Coulter).

Liang P., Henning S.M., Guan J., Grogan T., Elashoff D., Cohen P, & Aronson W.J. (2019). Effect of Dietary Omega-3 Fatty Acids on Castrate Resistant Prostate Cancer and Tumor Associated Macrophages. Prostate cancer and prostatic diseases, 23(1), 127-135.

+ Open protocol

+ Expand

Isolation and Characterization of Adipose Macrophages

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

It was performed after collagenase digestion and an additional wash with 10 mM EDTA to ensure the separation of lipid-laden macrophages, as we previously described (Nov et al. 2013 (link), Shapiro et al. 2013 (link)). After FcBlock (BD Biosciences, Franklin Lakes, NJ, USA), cells were stained with the following conjugated antibodies (10 min on ice in the darkness): CD45-APC, F4/80-PE-Cy7 (both from E-Bioscience, San Diego, CA, USA) and CD11b-APC-Cy7 (BD Pharmingen, San Diego, CA, USA). Cells were washed and pellets were then stained for 20 min on ice with BODIPY 493/503 (3 μg/mL BODIPY for 5 × 10⁶ cells; Invitrogen, D3922). Stained samples were further washed and filtered using 100 μm mesh. Propidium iodide (0.2 μg/mL; Sigma, P4864) was added to all samples. Stained samples were analyzed by FACS (Canto, BD Biosciences).

Vatarescu M., Bechor S., Haim Y., Pecht T., Tarnovscki T., Slutsky N., Nov O., Shapiro H., Shemesh A., Porgador A., Bashan N, & Rudich A. (2017). Adipose tissue supports normalization of macrophage and liver lipid handling in obesity reversal. The Journal of Endocrinology, 233(3), 293-305.

+ Open protocol

+ Expand

Multiparametric Flow Cytometry Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Erythrocytes-depleted spleen cells, MLN cells and PECs were analyzed using a 6-color FACS, following a standard protocol (34 (link)). Cells were stained with different combinations of conjugated monoclonal antibodies CD3-FITC (Biolegend, San Diego, CA, USA), CD19-PEcy7, CD8-APCcy7, Sca-1-PE, CD25-APC, CD69-PEcy7, CD86-PE (BD Biosciences), CD11b-APCcy7, CD11c-APC, CD4-PE, F4/80-PEcy7, MHC II-APC (eBioscience, San Diego, CA, USA), and CD40-FITC (Southern Biotech). In the first panel, cells were immunphenotyped using mAbs to CD3, CD19, CD11b, CD11c, and Sca-1. A second panel was used to analyze T cell subsets and their activation status and consisted of mAbs to CD3, CD4, CD8, CD25, and CD69. In the third panel, myeloid cell activation status was analyzed using mAbs to CD11b, F4/80, CD40, CD86, and MHC class II. Non-viable cells that stained positive for 7-AAD (eBioscience), were excluded from the analysis. For each antibody, appropriate isotype control was used. Data were collected on 30,000 cells using BD FACS Canto II (BD Biosciences) and analyzed using BD FACSDiva software (BD).

Al-Barazie R.M., Bashir G.H., Qureshi M.M., Mohamed Y.A., Al-Sbiei A., Tariq S., Lammers W.J., al-Ramadi B.K, & Fernandez-Cabezudo M.J. (2018). Cholinergic Activation Enhances Resistance to Oral Salmonella Infection by Modulating Innate Immune Defense Mechanisms at the Intestinal Barrier. Frontiers in Immunology, 9, 551.

+ Open protocol

+ Expand

Tracking Myeloid-Derived Suppressor Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

TRAMP-C1 cells (5×10⁵) were subcutaneously implanted into 6–8 week-old male C57BL/6 mice. Following the tumor formation, mice were sacrificed to sort Gr-1⁺, CD11b⁺, F4/80⁻ cells from the spleen when average volume of tumor reached 200mm³. Sorted cells, using Gr-1-APC eFluor 780, CD11b-eFluor 450, and F4/80-PE Cy7 antibodies (eBioscience, San Diego, CA), were labeled with CFSE (5 μM) for in vivo tracking. Labeled cells (1×10⁵ cells/mouse) were transferred into recipient male nude mice (6–8 week-old) bearing TRAMP-C1, TRAMP-C1^scram-sh, and TRAMP-C1^CRAMP-sh tumors by tail vein injection, when the average volume of tumors reached 800mm³. Recipient mice were sacrificed 3 days post-transfer for flow cytometry with the cells from the spleen and tumor.

Cha H.R., Lee J.H., Hensel J.A., Sawant A.B., Davis B.H., Lee C.M., Deshane J.S, & Ponnazhagan S. (2016). Prostate cancer-derived cathelicidin-related antimicrobial peptide facilitates macrophage differentiation and polarization of immature myeloid progenitors to protumorigenic macrophages. The Prostate, 76(7), 624-636.

+ Open protocol

+ Expand

Pulmonary Infiltrate Characterization During Pneumococcal Infection

Check if the same lab product or an alternative is used in the 5 most similar protocols

A single-cell suspension was generated from pulmonary digest to enable characterization of the pulmonary infiltrate during pneumococcal infection. A live/dead marker was first applied (LifeTechnologies L10119; 1:1,000) in PBS. Antibodies and blocker were made up in FACS buffer (PBS, 2% BSA, and 0.1% sodium azide): Fc receptors were blocked (1:100; anti-CD16/32, eBioscience 14-0161). Antibodies were incubated for 25 min at 4 °C. We used CD11b-PerCPCy5.5 (1:200; clone M1/70, 45-0112), CD11c-APC (1:400; clone N418, 17-0114), Ly6G-FITC (1:100; clone RB6-8C5, 11-5931), and F4/80-PE-Cy7 (1:200; clone BM8, 25-4801) (all eBioscience). After washing, cells were resuspended in 50 μL 1% PFA for 20 min at room temperature (RT). Cells were washed, resuspended in FACS buffer, and analysis was carried out on a BD FACS CANTO flow cytometer. Neutrophils were identified as f4/80⁻CD11b⁺Ly6G⁺. Alveolar macrophages were identified as F4/80⁺CD11c⁺CD11b^lo. Interstitial macrophages were identified as F4/80⁺CD11c⁻CD11b^hi.

Kitchen G.B., Cunningham P.S., Poolman T.M., Iqbal M., Maidstone R., Baxter M., Bagnall J., Begley N., Saer B., Hussell T., Matthews L.C., Dockrell D.H., Durrington H.J., Gibbs J.E., Blaikley J.F., Loudon A.S, & Ray D.W. (2020). The clock gene Bmal1 inhibits macrophage motility, phagocytosis, and impairs defense against pneumonia. Proceedings of the National Academy of Sciences of the United States of America, 117(3), 1543-1551.

+ Open protocol

+ Expand

Flow Cytometry Analysis of Tumor Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Tumor tissue was digested in a 0.3% collagenase/0.1% hyaluronidase solution, pressed through a nylon mesh filter to obtain a single cell suspension and incubated in red cell lysis buffer (0.17 M Tris-HCl, 0.16 M NH₄Cl) for 3 min, spun down, and resuspended in FACS buffer (PBS + 1.5% FBS). Equal numbers of cells were stained with a viability dye and combinations of the following antibodies: CD8a-PECy7 (eBioscience, San Diego, CA, USA, 25-0081-82), F4/80-PECy7(eBioscience, 25-4801-82), CD206-FITC (Biolegend, 141704), Ly6G-APC (eBioscience, 17-5931-81), CD11b-PE (eBioscience, 12-0112-82), CD45-PE-Cy5 (eBioscience, 15-0451-83), IFN-γ-APC (eBioscience, 17-7311-82), and SIINFEKL pentamer-PE (ProImmune, Oxford, UK). For autophagy analysis using Cyto-ID staining of bone marrow derived macrophages, cells were first stained for surface markers (CD80-PE-Cy5 (eBioscience, 15-0801-81) and CD206-APC (eBioscience, 17-2061-82)) followed by staining with Cyto-ID for 30 min at 37 °C, per the manufacturer’s protocol (Enzo Life Sciences, Farmingdale, NY, USA, ENZ-51031). Flow cytometric data were acquired on a BD FACSCanto II cytometer and analyzed using FACSDiva software version 9.2 (BD Biosciences, San Jose, CA, USA).

Alexander E.T., Fahey E., Phanstiel O I.V, & Gilmour S.K. (2023). Loss of Anti-Tumor Efficacy by Polyamine Blocking Therapy in GCN2 Null Mice. Biomedicines, 11(10), 2703.

+ Open protocol

+ Expand

Multi-color Flow Cytometry Immunophenotyping

Check if the same lab product or an alternative is used in the 5 most similar protocols

For direct multi-colour flow cytometry (LRS Fortessa X-20; BD Bioscience), cells were incubated for 30 min at 4 °C with the antibodies mentioned below. Live/dead cell discrimination was performed by staining with the Zombie Aqua Fixable Viability Kit (BioLegend, catalogue number: 423101). To block Fc receptors, a purified anti-mouse CD16/CD32 (eBioscience, catalogue number: 14-0161-86) was used.
CD19 APCCy7 (clone 6D5, Biolegend, catalogue number: 115530, 1:300), Ly6G PerCPCy5.5 (clone 1A8, Biolegend, catalogue number: 127616, 1:300), CD45 PEDazzle (clone 30-F11, Biolegend, catalogue number: 103146, 1:400), F4/80PECy7 (clone BM8, eBioscience, catalogue number:254801-82, 1:200), CD3 APC (clone 145-2c11, Biolegend, catalogue number: 100312, 1:100), CD11b Alexa700 (clone M170, Biolegend, catalogue number: 101222, 1:400), TCγδV421 (clone GL3, Biolegend, catalogue number: 118120, 1:100), CD4 BV605 (clone RM4-5, Biolegend, catalogue number: 100548, 1:300), CD8 BV711 (clone 53-6.7, Biolegend, catalogue number: 100748, 1:300) and NK1.1 PE (clone PK136, Biolegend, catalogue number: 557391, 1:300). IL-10^GFP production was measured in the FITC channel.

Mukherjee D., Chora Â.F., Lone J.C., Ramiro R.S., Blankenhaus B., Serre K., Ramirez M., Gordo I., Veldhoen M., Varga-Weisz P, & Mota M.M. (2022). Host lung microbiota promotes malaria-associated acute respiratory distress syndrome. Nature Communications, 13, 3747.

+ Open protocol

+ Expand

Characterization of Lung Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Resident cells were isolated from naive lung at CT0 and CT12 via lavage with BAL fluid. After staining with a live/dead marker (Life Technologies #L10119, 1:1000) in PBS, Fc receptors were blocked (1:100 anti-CD16/32, eBioscience #14-0161) before application of the following antibodies in 30 μl FACS buffer (PBS, 1% BSA and 0.1% sodium azide): CD11b-PerCP-Cy5.5 (1:200, clone M1/70, #45-0112), CD11c-APC (1:400, clone N418, #17-0114), Ly6G-FITC (1:100, clone RB6-8C5, #11-5931) and F4/80-PE-Cy7 (1:200, clone BM8, #25-4801) (all purchased from eBioscience). After washing, cells were resuspended in 50 μl FACS buffer and fixed by addition of an equal volume of 3.6% formaldehyde for 20 min. Cells were resuspended in FACS buffer, and analysis was carried out on a BD LSR II flow cytometer. Neutrophils were identified as F4/80⁻CD11b⁺Ly6G⁺. Alveolar macrophages were identified as F4/80⁺CD11c⁺CD11b^lo.

Gibbs J., Ince L., Matthews L., Mei J., Bell T., Yang N., Saer B., Begley N., Poolman T., Pariollaud M., Farrow S., Demayo F., Hussell T., Worthen G.S., Ray D, & Loudon A. (2014). An epithelial circadian clock controls pulmonary inflammation and glucocorticoid action. Nature medicine, 20(8), 919-926.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!