Anti f4 80 apc

Manufactured by BD

Sourced in United States

Anti-F4/80-APC is a fluorophore-conjugated antibody that binds to the F4/80 antigen, which is expressed on the surface of mouse macrophages. It is used in flow cytometry and other immunological applications to identify and quantify macrophage populations.

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Lab products found in correlation

Anti mhcii fitc, by BD (2 mentions) Anti cd86 pe, by BD (2 mentions) Cd64 bv421, by BioLegend (1 mentions) Anti cd40 pe, by BD (1 mentions) Anti cd45 v450, by BD (1 mentions) Anti ly6c pe cy7, by BD (1 mentions) Anti cd11b apc cy7, by BD (1 mentions) Ly6g percp cy5, by Thermo Fisher Scientific (1 mentions) Ccr2 apc, by Thermo Fisher Scientific (1 mentions) Cd11c pe txr, by R&D Systems (1 mentions) 70 m cell strainer, by BD (1 mentions) Collagenase type 4, by Merck Group (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Anti cd3 fitc, by BioLegend (1 mentions) Anti cd4 percp cy5, by BioLegend (1 mentions) Anti b220 percp cy5, by BioLegend (1 mentions) Anti dx5 fitc, by Thermo Fisher Scientific (1 mentions) Anti cd274 pe, by BD (1 mentions) Anti cd8 apc, by Thermo Fisher Scientific (1 mentions) Anti cd16 32 fc block, by BioLegend (1 mentions)

6 protocols using anti f4 80 apc

Monocyte and DC Recruitment in Mice

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To analyze monocyte and DC recruitment in response to CpG, 20 µl of a 100 µg/ml ODN1864 solution was injected into the footpad of C57BL/6 J mice. At the indicated time intervals post injection, blood samples were collected through the tail vein followed by ACK (Thermo Fisher Scientific) mediated red blood cell lysis. Alternatively, mice were euthanized and the DLNs were dissected. Single cell suspensions of DLN were prepared through incubation with collagenase type IV (Sigma-Aldrich) and passed through a 70 µm cell strainer (BD Falcon).
After treatment with 2.4G2 Ab for 5 min to block the FcR, PBMCs or lymph node cell suspensions were stained with the following anti-mouse Abs: anti-CD45-V450, anti- MHCII-FITC, anti-CD86-PE, anti-CD40- PE, anti-CD64-APC, anti-F4/80-APC, anti-Ly6C-PE-Cy7, anti-CD11b-APC-Cy7 (all BD Biosciences), Ly6G-PerCP-Cy5.5, CCR2-APC (eBioscience), CD64-BV421 (BioLegend) and CD11c-PE-TxR (R & D systems). Death cells were identified by staining with aqua life/dead stain (Invitrogen). Fluorescent events were acquired using an LSRII flow cytometer (BD Biosciences) and analyzed using FlowJo software. After exclusion of dead cells, granulocytes were identified as CD45+ CD11b+ Ly6C+ Ly6G+ cells and Monocytes as CD45+ Ly6C^hi CD11b^hi Ly6G- cells. Following exclusion of granulocytes and Monocytes in the LN, DCs were subdivided into MHCII^hi DCs and MHCII^int DCs.

De Koker S., Van Hoecke L., De Beuckelaer A., Roose K., Deswarte K., Willart M.A., Bogaert P., Naessens T., De Geest B.G., Saelens X., Lambrecht B.N, & Grooten J. (2017). Inflammatory monocytes regulate Th1 oriented immunity to CpG adjuvanted protein vaccines through production of IL-12. Scientific Reports, 7, 5986.

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Fluorescent Antibody Flow Cytometry

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The fluorescently labeled antibodies used for flow cytometry were purchased from the following companies, and the company protocols were followed in all applications: BioLegend (San Diego, CA, USA): anti-CD3-FITC (Cat. # 100306), anti-CD4-PerCP/Cy5.5 (Cat. # 100540), anti-B220-PerCP/Cy5.5 (Cat. # 103236), anti-CD3-PerCP/Cy5.5 (Cat. # 100328), anti-CD8-PerCP/Cy5.5 (Cat. # 100734), anti-CD8-APC (Cat. # 100712), and anti-CD279-APC (Cat. # 135209); eBioscience (San Diego, CA, USA): anti-DX5-FITC (Cat. # 11-5971-85), anti-CD274-PE (Cat. # 551892), and anti-F4/80-APC (Cat. # 17-4801-80); and BD Biosciences (San Jose, CA, USA): anti-CD 279-PE (Cat. # 551892).

Shindo Y., McDonough J.S., Chang K.C., Ramachandra M., Sasikumar P.G, & Hotchkiss R.S. (2016). Anti-Programed Cell Death Ligand 1 Peptide Improves Survival in Sepsis. The Journal of surgical research, 208, 33-39.

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Isolation of Splenic Dendritic Cells and Macrophages

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For isolation of splenic dendritic cells and macrophages, mouse spleens were perfused with 400 U/ml of collagenase D (Roche, Basel, Switzerland) in Hanks' balanced salt solution and incubated for 45 min at 37° followed by mechanical dissociation. Splenocytes were first incubated with anti‐CD16/32 (Fc‐block) (Biolegend, San Diego, CA) in PBS [1 mm EDTA, 2% fetal calf serum (FCS)] at 4° for 15 min, and were then stained with anti‐CD11c‐allophycocyanin (APC) (BD Biosciences, San Jose, CA) or anti‐F4/80‐APC (BD Biosciences, San Jose, CA) at 4° in PBS with 1 mm EDTA, 2% FCS. The BMDMs were first incubated with anti‐CD16/32 (Fc‐block) (Biolegend, San Diego, CA) in PBS (1 mm EDTA, 2% FCS) at 4° for 15 min. Cells were then stained with the following panel for 30 min at 4° in PBS (1 mm EDTA, 2% FCS): TLR2‐APC (Biolegend, San Diego, CA), CD206‐phycoerythrin/Cy7 (Biolegend, San Diego, CA), CD38‐BV510 (BD Biosciences, San Jose, CA) and F4/80‐APC/Cy7 (Biolegend, San Diego, CA). After washing twice, the cells were acquired using a Gallios flow cytometer (Beckman Coulter, Brea, CA) followed by data analysis using flowjo v10 (FlowJo, Ashland, OR).

Sjöstrand M., Carow B., Nyberg W.A., Covacu R., Rottenberg M.E, & Espinosa A. (2019). TRIM21 controls Toll‐like receptor 2 responses in bone‐marrow‐derived macrophages. Immunology, 159(3), 335-343.

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Investigating Tumor-Associated Macrophage Polarization in GL261 Glioma

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C57BL/6 mice were subcutaneously inoculated with GL261 cells (2 × 10⁶). When the tumor grew to 5 mm × 5 mm, the mice were randomly divided into two groups: NS and ARV-825 (20 mg/kg). After the mice were treated for three consecutive days, the tumor cells were obtained and stained with M2 type macrophage related antibodies (anti-CD45-PerCP-Cy5.5, anti-CD11b-PE, anti-F4/80-APC, anti-CD206-FITC) (BD Bioscience, USA) for 30 min at 4 °C, followed by washing and resuspending with PBS to perform FCM analysis.
The bone marrow derived macrophages (BMDMs) isolated from healthy female mice (6–8 weeks) were assigned to four groups (0, IL4, DMSO + IL4 and ARV-825 + IL4). Cells of DMSO + IL4 group and ARV-825 + IL4 group were respectively pretreated with DMSO and ARV-825 for 12 h. IL4 was then added to incubate for 24 h. The IL4 group and DMSO + IL4 group were the positive control groups and cells in 0 group were untreated. At the predetermined time point, cells were stained with mouse anti-CD45-PerCP-Cy5.5, anti-CD11b-FITC, anti-F4/80-PE and anti-CD206-APC antibodies (BD Bioscience, USA) for 30 min at 4 °C. Subsequently, stained cells were washed once and resuspended with PBS for FCM detection.

Yang T., Hu Y., Miao J., Chen J., Liu J., Cheng Y, & Gao X. (2022). A BRD4 PROTAC nanodrug for glioma therapy via the intervention of tumor cells proliferation, apoptosis and M2 macrophages polarization. Acta Pharmaceutica Sinica. B, 12(6), 2658-2671.

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Multiparametric Flow Cytometry Analysis

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Spleen and inguinal lymph node (LN) cells were harvested from immunized mice at the time of sacrifice. Joint tissues were minced and treated with collagenase (0.25 mg/ml), and joint cell populations were examined for surface markers using antibodies anti-CD4-AF700, anti-CD8a-PE-Cy7, anti-CD19-PE, anti-CD11b-Pacific Blue, anti-CD11c-APC-Cy7, anti-F4/80-APC, and anti-GR-1-FITC (BD Pharmingen, San Jose, CA, USA). For intracellular cytokine staining, cells were stimulated with Phorbol 12-myristate 13-acetate (25 ng/ml) and ionomycin (500 ng/ml) (Sigma-Aldrich) and treated with GolgiPlug (BD Pharmingen) for 4 hours. After cell surface staining with anti-CD3e-PE-Cy7 and anti-CD4-APC-Cy7, cells were permeabilized using the Cytofix/Cytoperm Plus kit (BD Pharmingen) and stained with anti-IFNγ-APC and anti-IL-17A-FITC. A BD LSRII cytometer was used for cytometry and data were analyzed using BD FACS Diva Software.

Olalekan S.A., Cao Y, & Finnegan A. (2014). Tissue specific CD4+ T cell priming determines the requirement for interleukin-23 in experimental arthritis. Arthritis Research & Therapy, 16(5), 440.

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Phenotypic Analysis of Peritoneal Macrophages

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Peritoneal macrophages obtained from control and treated mice were stained in a one-step test with the following fluorophore-labeled monoclonal anti-mouse mAbs: anti-F4/80-APC (BD Pharmingen, BM8), anti-CD11b-PerCP-Cy5.5 (BD Pharmingen, M1/70), anti-MHCII-FITC (BD Pharmingen, 25-9-17) and anti-CD86-PE (BD Pharmingen, GL-1). Phenotype analysis was carried out using the Becton Dickinson FACSCalibur apparatus with Cell Quest Software.

Kicielińska J., Szczygieł A., Rossowska J., Anger N., Kempińska K., Świtalska M., Kaszowska M., Wietrzyk J., Boratyński J, & Pajtasz-Piasecka E. (2016). Oral Administration of Polymyxin B Modulates the Activity of Lipooligosaccharide E. coli B against Lung Metastases in Murine Tumor Models. PLoS ONE, 11(2), e0148156.

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