48-2069-42

antibody from Invitrogen Antibodies

Targeting: MRC1 bA541I19.1, CD206, CLEC13D, CLEC13DL, MRC1L1

Provider product page for 48-2069-42

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Antibody data

Antibody Data
Antigen structure
References [16]
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Validations
Flow cytometry [1]
Other assay [19]

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Product number: 48-2069-42 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD206 (MMR) Monoclonal Antibody (19.2), eFluor™ 450, eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: This 19.2 monoclonal antibody reacts with human CD206, which is also known as the macrophage mannose receptor (MMR). CD206 is expressed on macrophages and dendritic cells. This type I transmembrane protein can also be detected on non-immune cells, including hepatic and lymphathic epithelia and kidney mesengial cells. CD206 binds to glycoproteins that terminate in D-mannose, L-fucose, or N-acetylglucosamine, as well as a variety of hormones. This receptor undergoes constitutive internalization and recycling between the plasma membrane and the endosomal compartment. CD206 is involved in antigen processing and presentation, cell migration, and intracellular signaling. Moreover, CD206 plays a key role in phagocytosis pathogens such as Candida albicans, Leishmania, and Mycobacterium tuberculosis. Applications Reported: This 19.2 antibody has been reported for use in flow cytometric analysis. Applications Tested: This 19.2 antibody has been pre-titrated and tested by flow cytometric analysis of GM-CSF-stimulated normal human peripheral blood cells. This can be used at 5 µL (0.25 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. eFluor® 450 is an alternative to Pacific Blue®. eFluor® 450 emits at 445 nm and is excited with the Violet laser (405 nm). Please make sure that your instrument is capable of detecting this fluorochome. Excitation: 405 nm; Emission: 445 nm; Laser: Violet Laser. Filtration: 0.2 µm post-manufacturing filtered.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: 19.2
Vial size: 100 Tests
Concentration: 5 µL/Test
Storage: 4° C, store in dark, DO NOT FREEZE!

MRC1 protein structure - 48-2069-42 shown in red.

Supportive validation

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Experimental details: NULL

Supportive validation

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Experimental details: NULL

Supportive validation

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Experimental details: Figure 2 miR-142-5p and miR-130a-3p regulate M2 activation of macrophages. ( a - d ) Human macrophages were transduced with nc, miR-142-5p ASO or miR-130a-3p mimics or both using lentiviral vectors. After 24 h, the cells were treated with IL-4 for 48 h. ( a ) The schematics of the approach. ( b ) Expression of CD206 and CD36 in macrophages as determined by flow-cytometry analysis. The representative histograms and quantitation of the MFI are shown (mean+-s.e.m., n =4 independent experiments; * P

Supportive validation

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Experimental details: Figure 9 Dyregulated Mph miR-142-5p and miR-130a-3p enhance liver fibrosis. ( a - f ) Mice were intravenously injected with LNA-modified miR-142-5p ASO, miR-130a-3p mimic or both every 3 days after CCL 4 challenge. Mice were sacrificed after 6 weeks (mean+-s.e.m., n =8 mice/group; * P

Supportive validation

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Experimental details: Figure 1 Metformin treated cancer cells polarized macrophage toward M1 phenotype THP-1 cells were stimulated with PMA (200 nM) for 24 h, then incubated with breast cancer (MDA-MB231/MDA-MB453) conditioned medium (CM) with or without metformin (60 muM) for 6 h, followed by flow cytometry analysis to quantify the amount of CD206, an M2 macrophage marker, and CD16, an M1 marker ( A, B ). Data are expressed as mean +- SD, * p < 0.05. DMSO: control; Met: metformin. Representative flow data shown are from experiments independently performed at least three times.

Supportive validation

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Experimental details: Figure 5 AMPK-NF-kappaB signaling participated in macrophage polarization Breast cancer cells (MDA-MB231) were treated with metformin 60 muM combined with an AMPK inhibitor (Compound C, CC) or NF-kappaB inhibitor (BAY-117082, BAY) for 6 h. The supernatant was collected to treat macrophages for 48 h, followed by flow cytometry analysis of CD206, M2 phenotype ( A ) and CD16, M1 phenotype ( B ). Data are expressed as mean +- SD, * p < 0.05. DMSO: control; CM: conditioned medium, Met: metformin. Representative flow data shown are from experiments independently performed at least three times.

Supportive validation

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Experimental details: 4 FIGURE M2-phenotype macrophages produce interleukin 6 (IL-6) in thyroid cancer. A, Flow cytometry showed that IL-6 was distributed in F4/80 staining-positive tumor-associated macrophages (TAMs). B, Flow cytometry showed that IL-6 was distributed in CD206-positive M2-phenotype macrophages. C, Double immunofluorescence analysis illustrated the colocalization of IL-6 and M2 phenotype within thyroid cancer

Supportive validation

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Experimental details: Figure 4 The polarizations of monocyte and GBM progression in a co-cultured model under hypoxic conditions. ( A ) THP-1 monocytes were co-cultured with U87-green fluorescent protein (GFP) or U251-GFP under hypoxic conditions for 48 h. The co-cultured cells were plotted on a side scatter versus FITC. The THP-1 monocytes (GFP-negative cells) were analyzed to assess the levels of cluster of differentiation (CD) 206, arginase 1 (Arg1), interleukin (IL)-10, interferon (IFN)-gamma, and tumor necrosis factor (TNF)-alpha by flow cytometry. ( B ) The median fluorescence intensity (MFI) of CD206, Arg1, IL-10, IFN-gamma, and TNF-alpha in THP-1 monocytes in the co-cultured model under hypoxic conditions for 48 h. * p < 0.05 compared with the THP-1 monocyte group. Quantitative data are presented as the mean +- standard error (representative of n = 3). ( C ) U87-GFP and U251-GFP were treated with U104 (CAIX inhibitor) and subsequently co-cultured with THP-1 monocytes under hypoxic conditions for 48 h. THP-1 monocytes (GFP-negative cells) were analyzed to assess the levels of CD206, Arg1, and IL-10 by flow cytometry. ( D ) U87-GFP or U251-GFP was treated with U104 and subsequently co-cultured with THP-1 monocytes under hypoxic conditions for 48 h. The co-cultured cells were plotted on a side scatter versus FITC. The GFP-positive gated GBM was analyzed to assess the levels of programmed death ligand 1 (PD-L1) by flow cytometry. ( E ) The levels of CAIX, PD-L1, tumor growth factor-beta, and c

Supportive validation

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Experimental details: Figure 2. The effect of TSN on macrophage polarization. (a) The comparisons of mRNA levels of cytokines from THP-1 stimulated by TSN of Panc-1-treated with electric fields. The mRNA levels of M1 markers increased along with the increasing electric fields, while the mRNA levels of M2 markers did not show a continuous increase along with the increasing electric fields. (b) The expression of surface markers of THP-1 stimulated by TSN of Bxpc-3 treated with electric fields. Increased expression of HLA-DR on THP-1 along with the increasing electric fields was observed. (c) IRE could increase infiltration of M1 macrophage in pancreatic tumor tissue (IHC). Orthotopic or subcutaneous tumor tissue was used as control. (d) IRE could increase the expression of CD16/32 in infiltrated macrophages in pancreatic tumor tissue. Orthotopic or subcutaneous tumor tissue was used as control. F4/80 was used to indicate macrophage; CD16/32 was the marker of M1 macrophage; CD206 was the marker of M2 macrophage; DAPI was used to indicate the nuclear of cells. (e) The expression of CD16/32 in macrophage from tumor tissue, spleen, peripheral blood, and lymph node in an orthotopic model of mouse pancreatic cancer after IRE treatment. IRE increased the expression of CD16/32 on the surface of macrophages from these tissues in the orthotopic model of mouse pancreatic cancer. (f) The expression of CD16/32 in macrophages from tumor tissue, spleen, peripheral blood, and lymph node in the subcutaneous model of

Supportive validation

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Experimental details: Fig. 3 Display of membrane skeleton-associated mannose patches is induced by oxidative stress and recognized by the mannose receptor on macrophages. a Peanut agglutinin (PNA) and Galanthus nivalis Agglutinin (GNA) lectin binding to normal hemoglobin (HbAA) RBCs with or without oxidation. Mannan blockade for GNA lectin binding shown in blue. 2 tailed Wilcoxon, paired data, PNA, n = 7; GNA, n = 8 biologically independent RBC donors over two independent experiments. b Immunofluorescence microscopy of GNA lectin/streptavidin (red) staining of healthy HbAA RBCs (above) and after the oxidative insult (below). c Normalized geometric mean fluorescence (gMFI) for binding analyzed by flow cytometry of murine Fc fusions with C-type lectins or sub-domains applied to oxidized versus undamaged RBCs. Mannan blockade of mannose receptor-carbohydrate recognition domain (MR-CRD) binding is shown in blue. MAH, macrophage antigen H. MR-CR, mannose receptor cysteine-rich domain. 2 tailed Wilcoxon, paired data, n = 7 biologically independent RBC donors over two independent experiments. d Immunofluorescence microscopy image of human monocyte-derived macrophages (HMDM) stained with DAPI (blue) and for mannose receptor (green) after incubation with oxidized HbAA RBCs, shown in magenta. e Percentage phagocytosis of oxidized RBCs by HMDM treated with human MR specific or scrambled siRNA. 2 tailed Mann-Whitney, n = 4 biologically independent RBC donors over two independent experiments. f Percentage phag

Supportive validation

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Experimental details: Fig. 4 Single-cell RNA sequencing of lung tissue from patients with severe COVID-19. ( A , C , and E ) Uniform Manifold Approximation and Projection (UMAP) plots showing individual populations of epithelial cells (A), macrophages (C), and mesenchymal cells (E). ( B , D , and F ) Heatmaps illustrating expression of select marker genes in epithelial cells (B), macrophages (D), and mesenchymal cells (F). Gene expression for the pulmonary fibrosis dataset of Habermann et al. () is shown as an average per condition; gene expression for the end-stage COVID-19 dataset is shown per individual patient. Labels on heatmaps (B, D, and F) correspond to the following samples: Control (H), healthy controls; IPF, idiopathic pulmonary fibrosis samples; Other PF, samples from patients with other forms of pulmonary fibrosis, all from the Habermann et al. dataset. Donor 1, Donor 2, control donor lungs; Case 1, lung transplant case 1; PMB1, PMB2, postmortem lung biopsies from two patients with COVID-19, all from the end-stage COVID-19 dataset. ( G ) Immunofluorescence microscopy revealed KRT17 staining (magenta) of flat epithelial cells resembling alveolar type 1 cells in nonfibrotic lung tissue from a patient who died of COVID-19 (palliative COVID-19). ( H and I ) Immunofluorescence microscopy revealed KRT17 staining (magenta) of distal explanted lung tissue from a patient with COVID-19 undergoing lung transplantation (H) and lung tissue from a patient (PMB1) who died from late-stage severe COVI

Supportive validation

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Experimental details: Figure 3 DRD2 and PD-L1 in GBM are involved in the polarization of TAMs in the GBM-macrophage co-cultured system. ( A ) BMDMs were co-cultured with ALTS1C1-GFP cells and then treated with or without PAL (20 muM) for 48 h. CD80 and CD206 expressions were determined by flow cytometry analysis. ( B ) Establishment of stable PD-L1 expression in ALTS1C1-GFP cells. The expression of PD-L1 was verified by Western blot. ( C ) BMDMs were co-cultured with PD-L1 ALTS1C1-GFP cells for 48 h. CD80 and CD206 were determined by flow cytometry analysis. ( D ) ALTS1C1-GFP cells were transfected with DRD2, and the expression of DRD2 was verified by real time-PCR. * p < 0.05 compared with shLacz group. ( E ) BMDMs were co-cultured with DRD2 knockdown of ALTS1C1-GFP cells for 48 h. CD80 and CD206 were determined by flow cytometry analysis.

Supportive validation

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Experimental details: Fig. 2 M2-TAM-derived exosomes enhance the stemness of GSCs. A TEM images of M2-TAM-derived exosomes. The presence of CD63, CD206, and CD163 in M2-TAM-derived exosomes analyzed by flow cytometric ( B ) and Western blot ( C ) analyses. D Fluorescence images of GSCs co-cultured with Dil-labeled M2-TAM-derived exosomes with nuclei stained with DAPI (blue). E Representative images showing neurosphere formation of GSCs along with the statistics of GSC formation rate and diameter of spheres. F Expression of stem cell-related protein CD133, Nestin, Oct4, Sox2, and GFAP in GSCs measured by Western blot analysis. G HE staining images of xenograft tumors from mice. H The representative immunohistochemical images of Sox2 in xenograft tumors and the percentage of GSCs labeled by Sox2. I Kaplan-Meier survive curve of tumor-bearing mice. n = 10. * p < 0.05 vs. the control EXO group. Measurement data were depicted as mean +- standard deviation, comparison of that between two groups was conducted by unpaired t test. Cell experiments were repeated three times independently.

Supportive validation

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Experimental details: FIGURE 2 Flow cytometry analysis of M1 and M2 surface markers (A) The expression of CD86 (M1 surface marker) and CD206 (M2 surface marker) on RAW264.7 cells were examined by flow cytometry. (B) Quantification of mean fluorescence intensity (MFI) of the surface markers ( n = 3, respectively). (C) The expression of CD86 and CD206 on THP-1 cells were examined by flow cytometry. (D) Quantification of mean fluorescence intensity (MFI) of the surface markers ( n = 3, respectively). The results are expressed as the mean values +- S.E.M. * p < 0.05, ** p < 0.01, *** p < 0.001.

Supportive validation

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Experimental details: Figure 1 Differential macrophage response to different inflammatory cytokine stimuli. ( A ) Morphological and immunocytochemical images showing monocyte-derived macrophage (MDM) polarization to M1 or M2 functional phenotype using type 1 cytokine IFN-gamma and lipopolysaccharide (LPS) or type 2 cytokine IL-4 and IL-13, respectively. ( B ) Increases in CD80+ M1 and CD206+ M2 MDMs were seen after treatment with IFN-gamma /LPS or IL-4/IL-13, respectively, using flow-cytometry analysis. Green arrows point to the CD80+ M1 macrophage, while red arrows indicate CD206+ M2 macrophages. ( C ) Flow cytometry and morphology imaging of M1 and M2 cell sorting and isolation using the fluorescence-activated cell sorting (FACS) assay. ( D ) Compared with the H1299 cells treated with M1 conditioned medium, the H1299 cells cultured with M2 conditioned medium exhibited higher invasion ability at 18 h in matrigel study. APC: antigen-presenting cell.

Supportive validation

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Experimental details: Figure 2 Astragalus polysaccharide (PG2) enhances M1 polarization and down-regulates IL-4/IL-13-induced M2 polarization. Images from flow cytometric analyses showing ( A ) the differentiation of THP-1 monocyte into macrophages after 24 h incubation in phorbol 12-myristate 13-acetate (PMA), ( B ) incubation of MDM in IL-4 and IL-13 induced a CD206 high CD80 low M2 phenotype, while ( C ) incubation of MDM in PG2 16 mg/mL for 48 h induced a CD80 high CD206 low M1 phenotype. ( D ) A graphical representation of the differential effect of IL-4/IL-13 and low dose (8 mg/mL) or high dose (16 mg/mL) PG2 treatment on M1-M2 polarization. ** p < 0.01

Supportive validation

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Experimental details: Figure 3 The enhancement of M1 macrophage polarization by PG2 is akin to the effect of LPS/IFN-gamma stimulation of MDMs. Images from flow cytometric analyses showing ( A ) the differentiation of THP-1 monocyte into macrophages after 24 h incubation in PMA; ( B ) MDMs after exposure to IFN-gamma and LPS induced a CD80 high CD206 low M1 phenotype; ( C ) cancer cell culture medium (CCCM) induced 17.1% CD80+ and 61.9% CD206+ MDMs; ( D ) PG2-treatment of MDMs pre-incubated in CCCM induced a CD80 high CD206 low M1 phenotype, similar to IFN-gamma/LPS exposure; ( E ) a graphical representation of the differential effect of IFN-gamma/LPS, CCCM, and 16 mg/mL PG2 treatment on M1-M2 polarization. PG2 enhanced the M1 phenotype akin to IFN-gamma/LPS exposure effect. ** p < 0.01

Supportive validation

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Experimental details: Figure 6 PG2 modulated the CD80+ M1/CD206+ M2 macrophage population and increased the population of CD80+, CD103+, and CD86+ dendritic cells derived from peripheral blood mononuclear cells (PBMCs) of cancer patient's ex vivo. The effect of PMA, LPS+INF-gamma, or PG2 on the proportion of ( A ) CD80+ and ( B ) CD206+ cells, as shown by flow cytometry. Graphical representation of the effect of PG2 on the population of ( C ) CD80+, ( D ) CD103+, and ( E ) CD86+ dendritic cells derived from GM-CSF+IL-4-treated PBMCs of NSCLC patients. ( F ) Graphical representation of the effect of PG2 on the number of functional CD103+ dendritic cells derived from GM-CSF+IL-4-treated PBMCs of breast, colon, ovarian, or liver patients. 1: GM-CSF+IL4; 2: GM-CSF+IL4, followed by the treatment of PG2 (16 mg/mL); 3: GM-CSF+IL4, washed out, and followed by the treatment of PG2 (16 mg/mL); * p < 0.05, ** p < 0.01.

Supportive validation

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Experimental details: Figure 3 Exogenous hrANXA1 controls human and mouse macrophage polarization in vitro. Human PBMC-derived macrophages were incubated for 24 hours with lipopolysaccharide plus IFN-gamma to promote an M1-like phenotype, before addition of human recombinant ANXA1 (hrANXA1; 10 nM) for a further 6 hours. ( A and B ) Median fluorescence intensity (MFI) units measured by flow cytometry of MHCII proinflammatory ( A ) and CD206 antiinflammatory ( B ) markers. Shown are MFI quantification (left) and representative FACS plots (right). ( C - F ) Quantitative reverse transcriptase PCR analysis of Tnfa ( C ) and Nos-2 ( D ) proinflammatory genes, and Il-10 ( E ) and Tgfb1 ( F ) antiinflammatory genes. ( G and H ) MFI units as measured by flow cytometry of MHCII proinflammatory ( G ) and CD206 antiinflammatory ( H ) markers after treatment by hrANXA1 in the presence or absence of the FPR2/ALX antagonist WRW 4 (10 muM). Experiments were performed on PBMCs from 6 ( A - F ) or 3 ( G and H ) independent donors. Each color represents an independent PBMC donor. * P < 0.05 vs. vehicle; # P < 0.05 vs. control.