The surface markers CCR7 (M1) and CD206 (M2) were examined via flow cytometry for evaluating different macrophage phenotypes. The specific procedure is the same as described in previous studies [5 (link)]. Anti-mouse CD16/32 (Biolegend, USA) was used to block the non-specific antigens. The antibodies for flow cytometry in this study included PE-conjugated CCR7 and PerCP-conjugated CD206 (Biolegend, USA), and the isotype controls were PE-conjugated Rat IgG2a, ĸ and PerCP-conjugated Rat IgG2a, ĸ (Biolegend, USA). The expression level of M1/M2 macrophage markers of all samples was analyzed using a flow cytometer (NovoCyte, ACEA Biosciences, USA) in triplicate. In addition, the surface markers IL-1β (M1) and CD163 (M2) were detected via western blot for further evaluating different macrophage phenotypes. The first antibodies for western blot were anti-IL-1β (ABclonal, Wuhan, China, the dilution was 1:1000), anti-CD163 (ABclonal, the dilution was 1:1000) and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (ABclonal, the dilution was 1:4000).
Pe conjugated ccr7
Manufactured by BioLegend
Sourced in United States
PE-conjugated CCR7 is a fluorochrome-labeled antibody that binds to the CCR7 chemokine receptor, which is expressed on various immune cell types. This product can be used in flow cytometry applications to identify and study cells expressing CCR7.
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Lab products found in correlation
3 protocols using pe conjugated ccr7
1
Macrophage Phenotype Evaluation Protocol
2
Macrophage Polarization on Titanium Surfaces
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BMDMs were seeded onto blank culture plates and Ti surface with different topographies in 24-well plates (1.5×105 cells/well). Macrophage surface markers CCR7 (M1) and CD206 (M2) were detected by flow cytometry for evaluating the different phenotypes. After 3-day incubation, cells were scraped off, centrifuged, and resuspended in purified anti-mouse CD16/32 (Biolegend, San Diego, CA, USA) for 10 min at 4°C to block the nonspecific antigens. Then, BMDMs were incubated with PE-conjugated CCR7 (Biolegend) and PerCP-conjugated CD206 (Biolegend) for 30 min at 4°C. The isotype controls were PE-conjugated rat IgG2a, κ and PerCP-conjugated Rat IgG2a, κ (Biolegend). After washing twice in PBS, BMDMs were resuspended in PBS and analyzed on a flow cytometer (NovoCyte; ACEA Biosciences, San Diego, CA, USA). All samples were analyzed in triplicate.
3
Quantifying Macrophage Subsets via Flow
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Briefly, RAW 264.7 cells were digested and blocked with CD16/32 for 5 min at 4°C. Then the cells were incubated with PE-conjugated CCR7 (BioLegend) and APC-conjugated CD206 (BioLegend) for 30 min at 4°C. After that, cells were analyzed by a flow cytometer (BD FACSCalibur). The data were analyzed by FlowJo software.
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