Anti mouse cd206

Manufactured by BD

Anti-mouse CD206 is a laboratory reagent used for the identification and quantification of the CD206 receptor in mouse samples. CD206, also known as the macrophage mannose receptor, is a cell surface glycoprotein expressed on the surface of macrophages and dendritic cells. This reagent can be used in various immunological techniques, such as flow cytometry and immunohistochemistry, to study the expression and distribution of the CD206 receptor in mouse models.

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5 protocols using anti mouse cd206

Visualizing Vascular Endothelial Cells in Mouse Tissues

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Mice were injected intravenous with 60 μg biotinylated lectin (Vector) and 60 μg streptavidin Cy3 (Invitrogen). After 5 minutes, mice were anesthetized and perfused with 20 mL of PBS and 20 mL of 3% paraformaldehyde injected into the left ventricle.
Tissues were fixed with 2% PFA for 5 to 6 hours, then washed with PBS overnight at 4°C Mesentery was fragmented into 0.2 to 0.5 cm and blocked in PBST (0.05% Tween20 in PBS) with 3% donkey serum for 30 minutes. Samples were then incubated with primary antibodies for 1 hour and washed with PBST, followed by secondary antibodies (1:200) for 1 hour. Antibodies used are antimouse CD31 (1:50; BD Biosciences), antimouse isolectin (1:75; Invitrogen), antimouse CD11b (1:50; BD Biosciences), and antimouse CD206 (1:50; BD Biosciences).

Moughon D.L., He H., Schokrpur S., Jiang Z.K., Yaqoob M., David J., Lin C., Iruela-Arispe M.L., Dorigo O, & Wu L. (2015). Macrophage Blockade Using CSF1R Inhibitors Reverses the Vascular Leakage Underlying Malignant Ascites in Late-Stage Epithelial Ovarian Cancer. Cancer research, 75(22), 4742-4752.

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Tumor Immunotherapy Protocol

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Anti-mouse Tim-3 (clone: RMT3-23), anti-mouse PD-1 (clone: J43), anti-mouse Lag-3 (clone: C9B7W), Rat IgG isotype control (clone: 2A3) and polyclonal American hamster IgG were purchased from Bioxcell company for tumor therapy. For Flow cytometry, anti-mouse CD45 (clone: 30-F11), anti-mouse CD4 (clone: GK1.5), anti-mouse CD8a (clone: 53–6.7), anti-mouse Foxp3 (MF23), anti-mouse γδTCR (clone: GL3), anti-mouse CD103 (clone: M290), anti-mouse Tim-3 (clone: 5D12), anti-mouse Lag-3 (clone: C9B7W), anti-mouse Lag-3 (clone: C9B7W), anti-mouse CD206 (clone: MR5D3), anti-mouse CD62L (clone: MEL-14), anti-mouse CD44(clone: IM7), anti-mouse OX40 (OX-86), anti-mouse IL7R (clone: SB/199), anti-mouse GITR (clone:DTA-1) and anti-mouse CD11b (clone: M1/70) were purchased from BD Bioscience. anti-mouse Ki67(clone: SolA15), IFN-γ (clone: XMG1.2), CD11b (M1/70) were purchased from ebioscience. anti-mouse B220 (clone: RA3-6B2), anti-mouse CD8a (clone: 53–6.7), anti-human/mouse Granzyme B (clone: GB11), anti-mouse Granzyme A (clone: 3G8.5), anti-mouse MHC II (clone: M5/114.15.2), anti-mouse Gr-1 (clone: RB6-8C5), anti-mouse CD24 (clone: M1/69), anti-mouse F4/80 (clone: BM8) and anti-mouse NK1.1 (clone: PK136) were purchased from Biolegend. Mitotracker Deep Red FM labeling kit was purchased from Invitrogen. Ghost 510 and Zombie NIR dye was purchased from Tonbo Biosciences and Biolegend.

Yang M., Du W., Yi L., Wu S., He C., Zhai W., Yue C., Sun R., Menk A.V., Delgoffe G.M., Jiang J, & Lu B. (2020). Checkpoint molecules coordinately restrain hyperactivated effector T cells in the tumor microenvironment. Oncoimmunology, 9(1), 1708064.

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Immune Cell Profiling in Small Intestine

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After blocking Fc receptors with anti-mouse CD16/CD32 (BD Biosciences), small intestinal epithelial cells were stained with anti-mouse Ly-6G (BioLegend), anti-mouse F4/80 (BD Biosciences), anti-mouse CD86 (BD Biosciences), anti-mouse CD206 (BD Biosciences), and anti-mouse CD45 (BD Biosciences). The stained cells were analyzed on a FACSCalibur flow cytometer (BD Biosciences). The data were analyzed using FlowJo Software (FlowJo, Ashland, OR). Neutrophils were defined as Ly6G⁺ cells and macrophages as F4/80⁺ cells and M1 macrophages as F4/80⁺ CD86⁺ and M2 macrophages as F4/80⁺ CD206⁺. Lymphocytes were defined as CD45⁺ cells.

Ho J., Chan H., Liang Y., Liu X., Zhang L., Li Q., Zhang Y., Zeng J., Ugwu F.N., Ho I.H., Hu W., Yau J.C., Wong S.H., Wong W.T., Ling L., Cho C.H., Gallo R.L., Gin T., Tse G., Yu J., Chan M.T., Leung C.C, & Wu W.K. (2020). Cathelicidin preserves intestinal barrier function in polymicrobial sepsis. Critical Care, 24, 47.

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Mouse Bone Cell Isolation for Flow Cytometry

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For flow cytometry analysis, the mouse femur was cut into pieces, digested in 1 mg/ml collagenase I, 1 mg/ml dispase II at 37°C for 30 min, centrifuged. After lysis of red blood cells on ice, the samples were passed through a 70-μm filter, centrifuged, and ready for staining. Anti-mouse CD45, anti-mouse CD11b, anti-mouse CD86, and anti-mouse CD206 were purchased from BD Biosciences, and the permeabilization/fixation kit was purchased from eBioscience. All staining processes were performed in 100 μl PBS. The cells were stained on ice for 30 min and then analyzed by flow cytometry (BD Biosciences).

Chen M., Lin W., Ye R., Yi J, & Zhao Z. (2021). PPARβ/δ Agonist Alleviates Diabetic Osteoporosis via Regulating M1/M2 Macrophage Polarization. Frontiers in Cell and Developmental Biology, 9, 753194.

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Quantifying Lung Macrophages in Angpt1 Mice

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Macrophages in the lung were evaluated by FACS 3 weeks after tail vein injection of 1 × 10⁵ B16F10 cell in Angpt1^Δ/Δ and WT mice. Lungs were dissected and digested into single cell suspension by incubation in Hank’s Balanced Salt Solution (HBSS) containing 1 mg/ml Collagenase I, 1 mg/ml Collagenase IV, 1 mg/ml Collagenase V, and 1 U/ml DNAse I for 45 min at 37°C. The cells were then washed 3 times by centrifugation at 500 x g for 5 min and exchange of buffer, HBSS with 3% BSA and 1 mM EDTA. Lung suspensions were stained with anti-mouse CD36 (BD Bioscience) for macrophages and anti-mouse CD206 (BD Bioscience) to identify changes in macrophage polarization between Angpt1^Δ/Δ and WT mice in a BD FACS Calibur. In addition, staining with anti-Tek (124,007, Biolegend) was done. Mean fluorescence intensity (MFI) was calculated using BD FlowJo software (BD Bioscience). Quantification of macrophages labelled with Isolectin B4 was done on vibratome sections of lung using a Leica SP8 confocal microscope.

Michael I.P., Orebrand M., Lima M., Pereira B., Volpert O., Quaggin S.E, & Jeansson M. (2017). Angiopoietin-1 deficiency increases tumor metastasis in mice. BMC Cancer, 17, 539.

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