rat anti mouse cd206 fitc  (Bioss)

Journal: Frontiers in Immunology

Article Title: Bone Marrow-Derived and Elicited Peritoneal Macrophages Are Not Created Equal: The Questions Asked Dictate the Cell Type Used

doi: 10.3389/fimmu.2020.00269

Figure Lengend Snippet: Comparison of mRNA expression from BMDM and pMACs in response to polarization.

Article Snippet: CD206 , Mannose Receptor , CD206-FITC , MR5D3 , AbD Serotec , — , — , .

Techniques: Comparison, Expressing

Journal: Frontiers in Immunology

Article Title: Bone Marrow-Derived and Elicited Peritoneal Macrophages Are Not Created Equal: The Questions Asked Dictate the Cell Type Used

doi: 10.3389/fimmu.2020.00269

Figure Lengend Snippet: List of Antigens, genes, and released proteins used in these studies.

Article Snippet: CD206 , Mannose Receptor , CD206-FITC , MR5D3 , AbD Serotec , — , — , .

Techniques: Multiplex Assay

Journal: Scientific Reports

Article Title: Effects of scavenger receptors-1 class A stimulation on macrophage morphology and highly modified advanced glycation end product-protein phagocytosis

doi: 10.1038/s41598-018-24325-y

Figure Lengend Snippet: AGE-2 and AGE-3 effects on scavenger receptor expression. ( a ) RAW264.7 cells seeded at 2 × 10 5 per well incubated for 1 h with 200 μg/ml AGE-2, AGE-3, or BSA were stained with specific FITC-labelled Abs against CD163 or CD206, specific PE-labelled CD204 or LOX-1, and APC-labelled CD36 or RAGE, and relative mean fluorescence intensities (MFIs) determined by flow cytometry (n = 6). Each column represents respective surface protein MFIs relative to medium only (Untreated, Unt) after 1 h incubation, arbitrarily defined as 100%. Data are presented as the means ± SEM and analysed using one-way ANOVA followed by Tukey’s test. *** p < 0.001, ** p < 0.01 compared with the Unt value. ( b ) For RAW264.7 cells seeded at 2 × 10 5 per well incubated with increasing concentrations AGE-2, AGE-3, and BSA (0.2–200 μg/ml) for 1 or 4 h, relative CD204 MFIs were determined by flow cytometry (n = 4–6). Each column represents MFIs relative to Unt after 1 or 4 h incubation, arbitrarily defined as 100%. Data are presented as the means ± SEM and analysed using one-way ANOVA followed by Tukey’s test. *** p < 0.001, * p < 0.05 compared with Unt. Insert in 1 h shows overlay of representative monoparametric histogram of CD204 expression in RAW264.7 cells incubated with medium only (Untreated; greyscale), 200 μg/ml BSA (orange), AGE-2 (blue), or AGE-3 (red line). ( c ) RAW264.7 cells seeded at 4 × 10 5 per well were cultured for 4 h with 200 μg/ml Alexa Fluor 488 (green)-labelled BSA, AGE-2, or AGE-3. CD204 intracellular localisation was visualized using anti-CD204 Ab and Alexa Fluor 594 (red) labelled secondary Ab, with confocal fluorescence microscopy. Rabbit IgG served as an anti-CD204 Ab isotype-matched control. All micrographs are shown at the same magnification (×400). Scale bar indicates 50 μm. Each BSA, AGE-2, or AGE-3 panel indicates, as follows; (upper) Alexa Fluor 488-labelled AGEs, (middle) Alexa Fluor 594-labelled CD204, or (lower) merged image. Clipped images at the upper left of the merged panel in AGE-2 or AGE-3 represent higher magnifications of the white rectangle region in the corresponding panel. Scale bar indicates 10 μm.

Article Snippet: Subsequently, cells were harvested and rinsed with FACS wash buffer followed by centrifugation (200 × g , 5 min, 4 °C), then stained with anti-mouse Abs against phycoerythrin (PE)-conjugated CD204 (4 ng, 130-102-328, Miltenyi Biotec, Bergisch Gladbach, Germany), PE-conjugated TLR4 (50 ng, 12-9041-80, Thermo Fisher Scientific), PE-conjugated LOX-1 (0.5 μl, FAB1564P, R&D Systems), allophycocyanin (APC)-conjugated CD36 (25 ng, 102-612, BioLegend, San Diego, CA, USA), APC-conjugated RAGE (50 ng, LS-C212626, LifeSpan BioSciences, Seattle, WA, USA), fluorescein isothiocyanate (FITC)-conjugated CD163 (200 ng, bs-2527R-FITC, Bioss, Woburn, MA, USA), or FITC-conjugated CD206 (200 ng, MCA2235F, Bio-Rad Laboratories, Berkeley, CA, USA) at 4 °C for 30 min. After rinsing with FACS wash buffer followed by centrifugation (200 × g , 5 min, 4 °C), 300 μl of PBS (−) was added to the residue followed by staining with PI (2 μg/ml), FACS CantoII analysis, and data processing using BD FACSDiva software to determine the MFI of each surface membrane receptor.

Techniques: Expressing, Incubation, Staining, Fluorescence, Flow Cytometry, Cell Culture, Microscopy, Control

Journal: Scientific Reports

Article Title: Effects of scavenger receptors-1 class A stimulation on macrophage morphology and highly modified advanced glycation end product-protein phagocytosis

doi: 10.1038/s41598-018-24325-y

Figure Lengend Snippet: Inhibitory effect of Abs against scavenger receptors for AGE uptake RAW264.7 cells seeded at 2 × 10 5 per well were pre-incubated with or without neutralizing Abs against scavenger receptors at 20 or 10 μg/ml for 1 h followed by culturing with BSA, AGE-2, or AGE-3 at 200 μg/ml for 1 h. ( a ) Neutralizing effect of anti-CD204 Ab on the uptake of fluorescent AGE-2, AGE-3, and BSA was determined by flow cytometry (n = 3). Goat IgG was used as an isotype-matched control of anti-CD204 Ab. ( b ) Neutralizing effect of each Ab against RAGE, LOX-1, CD36, CD163, or CD206 on the uptake of fluorescent BSA, AGE-2, or AGE-3 was determined by flow cytometry (n = 3–10). ( c ) Effect of anti-CD204 Ab on the expression of RAGE, LOX-1, CD36, CD163, and CD206 was determined by flow cytometry (n = 4–5). Each column represents the MFIs of the respective surface membrane proteins relative to the medium only group (Unt, Untreated) after 1 h incubation, which was arbitrarily defined as 100%. Data are presented as the means ± SEM and were analysed using one-way ANOVA followed by Tukey’s test. *** p < 0.001, ** p < 0.01 compared with the value for Unt. ### p < 0.001, ## p < 0.01 compared with the value for AGE-2 alone. ††† p < 0.001 compared with the value for AGE-3 alone.

Article Snippet: Subsequently, cells were harvested and rinsed with FACS wash buffer followed by centrifugation (200 × g , 5 min, 4 °C), then stained with anti-mouse Abs against phycoerythrin (PE)-conjugated CD204 (4 ng, 130-102-328, Miltenyi Biotec, Bergisch Gladbach, Germany), PE-conjugated TLR4 (50 ng, 12-9041-80, Thermo Fisher Scientific), PE-conjugated LOX-1 (0.5 μl, FAB1564P, R&D Systems), allophycocyanin (APC)-conjugated CD36 (25 ng, 102-612, BioLegend, San Diego, CA, USA), APC-conjugated RAGE (50 ng, LS-C212626, LifeSpan BioSciences, Seattle, WA, USA), fluorescein isothiocyanate (FITC)-conjugated CD163 (200 ng, bs-2527R-FITC, Bioss, Woburn, MA, USA), or FITC-conjugated CD206 (200 ng, MCA2235F, Bio-Rad Laboratories, Berkeley, CA, USA) at 4 °C for 30 min. After rinsing with FACS wash buffer followed by centrifugation (200 × g , 5 min, 4 °C), 300 μl of PBS (−) was added to the residue followed by staining with PI (2 μg/ml), FACS CantoII analysis, and data processing using BD FACSDiva software to determine the MFI of each surface membrane receptor.

Techniques: Incubation, Flow Cytometry, Control, Expressing, Membrane