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Ccr5 apc

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The CCR5-APC is a fluorescently labeled antibody that binds to the CCR5 receptor, a chemokine receptor expressed on the surface of various immune cells. This product is designed for use in flow cytometry applications to identify and analyze cells expressing the CCR5 receptor.

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Lab products found in correlation

Cx3cr1 fitc, by BioLegend (2 mentions) Fortessa 2, by BD (1 mentions) Cd3 apc efluor780, by Thermo Fisher Scientific (1 mentions) Cd19 apc efluor780, by Thermo Fisher Scientific (1 mentions) Fixable viability dye efluor 450, by Thermo Fisher Scientific (1 mentions) Cd45 pe, by Thermo Fisher Scientific (1 mentions) Perforin apc, by BioLegend (1 mentions) Granzyme b fitc, by BioLegend (1 mentions) Cd16 fitc, by Thermo Fisher Scientific (1 mentions) Cd69 apc, by BioLegend (1 mentions) Cd103 fitc, by BioLegend (1 mentions) Human foxp3 buffer, by BD (1 mentions) Cxcr6 percp cy5, by BioLegend (1 mentions) Hla a2 fitc, by BioLegend (1 mentions) Gm csf pe, by BioLegend (1 mentions) Cd94 fitc, by Thermo Fisher Scientific (1 mentions) Cxcr6 apc, by BioLegend (1 mentions) T bet pecy7, by Thermo Fisher Scientific (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Cd45 fitc, by Thermo Fisher Scientific (1 mentions)

5 protocols using ccr5 apc

1

Differential Immunophenotyping of NK Cell Subsets

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The following antibodies were used. From eBioscience (San Diego, CA): CD3-APC eFluor 780 (clone SK7); CD16-FITC (eBioCB16); CD19-APC eFluor 780 (HIB19); CD45-PE (HI30); CD94-FITC (DX22); Eomes-PE eFluor 610 (WD1928); Granzyme K-PerCP eFluor 710 (G3H69); HLA-A3-FITC (GAP.A3); IFN gamma-Alexa Fluor 488 (4S.B3); S1PR1-eFluor 660 (SW4GYPP); T-bet-PECy7 (4B10); TNF alpha-APC (Mab11). From Biolegend (London, UK): CCR5-APC (J418F1); CD49a-FITC (TS2/7); CD69-APC (FN50); CD103-FITC (Ber-ACT8); CX3CR1-FITC (2A9-1); CXCR6-PerCP Cy5.5 (K041E5); CXCR6-APC (K041E5); GM-CSF-PE (BDV-21C11); Granzyme B-FITC (GB11); HLA-A2-FITC (BB7.2); KIR2DL1/S1/S3/S5-APC (HP-MA4); KIR2DL2/L3-APC (DX27); KIR3DL1-APC (DX9); Perforin-APC (dG9). From BD (Oxford, UK): CD56-BV510 (NCAM16.2), LIF-PE (1F10). Dead cells were excluded using Fixable Viability Dye eFluor 450 (eBioscience). Intracellular staining was carried out using Human FoxP3 Buffer (BD) according to the manufacturer’s instructions. Data were acquired on a Fortessa II (BD) and analysed using FlowJo (Treestar, Ashland, OR). Cells were sorted on an Aria (BD). Eomeslo NK cells were isolated by sorting on live cells (propidium iodide negative, Tonbo Biosciences, San Diego, CA), singlets, scatter, CD3- CD56+ CXCR6- CD16+. Eomeshi NK cells were isolated by sorting on live cells, singlets, scatter, CD3- CD56+ CXCR6+.
Cuff A.O., Robertson F.P., Stegmann K.A., Pallett L.J., Maini M.K., Davidson B.R, & Male V. (2016). Eomeshi NK Cells in Human Liver Are Long-Lived and Do Not Recirculate but Can Be Replenished from the Circulation. The Journal of Immunology Author Choice, 197(11), 4283-4291.
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2

Flow Cytometry Analysis of Immune Cell Subsets

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Flow cytometry was performed as described (6 (link)). Cells were incubated with anti-mouse CD16/32 (Fc Block; BD Biosciences) before staining with primary antibody or isotype controls. Ten-thousand to 50,000 events per sample were acquired using an LSRII flow cytometer (BD-Biosciences) and analyzed with Flowjo flow cytometry software (Tree Star Inc.). The following antibodies were used: CD11b-Brilliant Violet 421, Tim4-PE, CD138-APC, CD138-PE F4/80- Pacific Blue, Ly6G-APC-Cy7, Ly6C-APC-Cy7, CD80-PerCp-Cy5, CD40-PerCP-Cy5, CD206- PerCP-Cy5, CCR2-FITC, CX3CR1-FITC, CD169-APC, CCR5-APC, CD36-PE, CD36-AlexaFluor 488, and IL-10R-PE (Biolegend); Marco-FITC (Biorad); Ly6C-FITC, CD86-FITC, CD11c-FITC, I-A/I-E-PE, CD93-PE (BD Bioscience); CREB-PE and Phospho-CREB-FITC (Cell Signaling); CD11b-Pacific Blue, CD45-FITC, TLR4-PE, CD45-FITC (eBioscience). Buffers used in intracellular staining were from eBioscience. For cell-sorting, PEC from untreated B6 mice were stained with anti-CD11b, Tim4, and CD138 antibodies. CD11b+Tim4+ and CD11b+CD138+ cells were gated and sorted (FACSaria cell sorter). Forty-thousand cells/subset were collected and lysed immediately for RNA extraction.
Han S., Zhuang H., Shumyak S., Wu J., Li H., Yang L.J, & Reeves W.H. (2017). A novel subset of anti-inflammatory CD138+ macrophages is deficient in mice with experimental lupus*. Journal of immunology (Baltimore, Md. : 1950), 199(4), 1261-1274.
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3

Flow Cytometry Characterization of TSLF Cells

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TSLF cells were stained with CD4-PE-Cy7 (BD, USA), CCR5-APC (BioLegend, USA), and CD184-fluorescein isothiocyanate (FITC) (BioLegend, USA) to detect CD4, CCR5, and CXCR4 expression, respectively. CXCR4-PECy7 (BioLegend, USA) was used to detect CXCR4 knockdown. The percentage of GFP-positive cells was examined on a BD Biosciences FACSVerse flow cytometer (USA) driven by FACSuite v1.0.3. Analysis of the acquired data was performed using FlowJo v7.6.1 (TreeStar Inc., USA).
In the flow cell-sorting experiment, mCherry-positive cells were sorted using a Sony SH800 flow cytometer (Japan) driven to gain TSLF-CD4 tsA3Z2c-Z1b knockout cells.
Luo M.T., Mu D., Yang X., Luo R.H., Zheng H.Y., Chen M., Guo Y.Q, & Zheng Y.T. (2021). Tree Shrew Cells Transduced with Human CD4 and CCR5 Support Early Steps of HIV-1 Replication, but Viral Infectivity Is Restricted by APOBEC3. Journal of Virology, 95(16), e00020-21.
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4

Multiparametric Immune Cell Profiling

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Cell staining was performed using fluorescence-conjugated antibodies, specifically CD1a-Alexa Fluor 488, CD14-PE, CD11c-APC, CD1c-FITC, CD16-APC, CD86-Alexa Fluor 488, CD83-PE, CD80-PerCP/Cy5.5, CD40-APC, human leucocyte antigen (HLA)-DR-Alexa Fluor 488, HLA-ABC-APC, CCR C-C chemokine receptor 1 (CCR1)-Alexa Fluor 488, CCR2-PerCP/Cy5.5, CCR5-APC, CCR7-PerCP/Cy5.5, chemokine receptor (CXCR4)-PE, CD3-PE, CD4-PerCP/Cy5.5, CD8-APC, CD69-FITC, CD25-APC, forkhead-box-P3 (FoxP3)-FITC, T-box protein expressed in T-cells (T-bet)-PE (all from Biolegend). Isotype-matched antibodies were used as controls. Briefly, 0.2 × 106 monocytes, DCs or T cells were washed and stained with 3 µl of fluorescence-conjugated antibodies in phosphate-buffered saline (PBS) + 1% FBS for 30 min at 4°C, in the dark. Cells were subsequently washed, resuspended in PBS + 1% FBS and analyzed in an Accuri C6 flow cytometer (BD Bioscience, San Jose, CA, USA). For intracellular staining, Fix&Perm (Thermo FisherScientific, Waltham, MA, USA), a fixation and cell permeabilization kit, was used as described by the manufacturer. Data were analyzed with FlowJo™ software (version 10) and results presented as percentage of positive cells or mean fluorescence intensity (MFI) after subtraction of isotype control values.
Calmeiro J., Mendes L., Duarte I.F., Leitão C., Tavares A.R., Ferreira D.A., Gomes C., Serra J., Falcão A., Cruz M.T., Carrascal M.A, & Neves B.M. (2021). In-Depth Analysis of the Impact of Different Serum-Free Media on the Production of Clinical Grade Dendritic Cells for Cancer Immunotherapy. Frontiers in Immunology, 11, 593363.
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5

Monocyte Phenotyping by Flow Cytometry

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All flow cytometry tests were performed on freshly prepared PBMCs and CD14 þ CD16 -monocytes. After blocking nonspecific antibody binding with human TruStain FcX (BioLegend, USA), the following monoclonal antibodies were used for flow cytometry: CD14-FITC, CD16-BV421, CCR1-PE, CCR2-PE/Cy7, CCR5-APC, CD11b-APC, CD18-PE (all from BioLegend, USA). We performed compensation using CompBeads (Becton Dickinson, USA). Flow cytometric analysis was performed on the CytoFLEX platform (Beckman Coulter, USA). The acquired data were analyzed by FlowJo software (Becton Dickinson, USA). Unstained monocytes were used as a negative control. Monocytes in PBMCs were identified as HLA-DR þ CD3 À CD19 -population with the unique forward and side-scatter properties.
Zhao X., Gu M., Xu X., Wen X., Yang G., Li L., Sheng P, & Meng F. (2020). CCL3/CCR1 mediates CD14+CD16- circulating monocyte recruitment in knee osteoarthritis progression. Osteoarthritis and cartilage, 28(5).
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