Cultured T cells were washed in PBS/0.1% BSA and re-suspended at 1×106 cells in 50 uL Brilliant Buffer (BD Biosciences) supplemented with 4% rat serum for 15 minutes at 4C. Cells were then incubated for 30 minutes at 4°C in 100 μL of Brilliant Buffer using the following antibody fluorophore conjugates (all from BD Biosciences unless otherwise noted): CD7 BV421, CD4 BV510, CCR4 BV605 (BioLegend), CD8 BV650, CD196 BV786 (BioLegend), CD3 AF488, CD45RA PerCPCy5.5, CD183 PE, CD197 PE-CF594, CD185 PE-Cy7 (BioLegend), and CCR10 APC (R&D Systems). Full details of fluorophore conjugated antibodies can be found in supplemental table 5 . Cells were then washed twice in PBS/0.1% BSA and data acquired on a ZE5 (Yeti) cytometer (BioRad/Propel Labs). Compensation and analyses were performed on FlowJo V10 (TreeStar) using fluorescence minus one (FMO) controls. Statistical analyses were performed on GraphPad Prism 7 using 2-way ANOVA with Bonferroni post-hoc corrections.
Ccr10 apc
Manufactured by R&D Systems
The CCR10 APC is a lab equipment product manufactured by R&D Systems. It is a fluorochrome-conjugated monoclonal antibody that binds to the human CCR10 receptor, which is expressed on the surface of certain cell types. The CCR10 APC can be used for the identification and analysis of CCR10-expressing cells in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Brilliant buffer, by BD (2 mentions)
Cd4 bv510, by BioLegend (2 mentions)
Ccr4 bv605, by BioLegend (2 mentions)
Cd8 bv650, by BioLegend (2 mentions)
Cd3 af488, by BioLegend (2 mentions)
Cd45ra percp cy5, by BioLegend (2 mentions)
Cd183 pe, by BioLegend (2 mentions)
Cd7 bv421, by BioLegend (2 mentions)
Cd196 bv786, by BioLegend (2 mentions)
Cd197 pe cf594, by BioLegend (2 mentions)
Cd185 pe cy7, by BioLegend (2 mentions)
Cd4 pe cy7, by BioLegend (2 mentions)
Cd69 apc, by BioLegend (2 mentions)
Cd69 bv421, by BioLegend (2 mentions)
Hla dr apc, by BioLegend (2 mentions)
Mouse fitc cd4 and efluor 605 cd44, by BD (2 mentions)
Brilliant violet 570 cd4, by BioLegend (2 mentions)
Alexa fluor 700 cd44, by BioLegend (2 mentions)
Fitc klotho, by Bioss Antibodies (2 mentions)
Pe cy7 cd99, by Bioss Antibodies (2 mentions)
5 protocols using ccr10 apc
1
Multiparameter Flow Cytometry of T Cells
2
Lymphocyte Isolation and Characterization
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Lymphocytes were isolated as described and stained with anti-mouse IgD PerCP-Cy5.5 (1:400, 405710), LPAM (integrin α4β7) PE (1:100, 120605; (Biolegend, San Diego, CA), B220 V500 (1:400, 561227), CD19 APC-H7 (1:200, 560143), CD80 PE (1:500, 553769), CD273 APC (1:200, 560086), CD138 PE (1:200, 553714), IgM PE-Cy7 (1:200, 552867; BD Biosciences), CCR9 PE-Cy7 (1:100, 25-1991), CD73 PE-Cy7 (1:50, 25-0731), IgA PE (1:50, 12-4204), GL7 eFluor 450 (1:100, 48-5902), CD38 Alexa700 (1:800, 56-0381), CD21/35 Pacific Blue (1:800, 57-0212; eBiosciences) or CCR10 APC (1:100, FAB2815A; R and D systems. Minneapolis, MN) and were analysed using LSR II (BD Biosciences) or Navios (Beckman Coulter, Brea, CA) flow cytometers. For sorting, cells were labelled with anti-mouse CD138 PE (1:200, 553714), CD19 PE-Cy7 (1:200, 552854), CD80 APC (1:200, 560016) and GL7 FITC (1:100, 553666) before sorting using a FACSAria (BD Biosciences). Cells were sorted into tubes that had been coated with 2% BSA/PBS overnight, and pelleted by centrifugation at 600 g before being resuspended in PBS and injected into recipient mice or used for gene sequence analysis.
3
Multiparameter Flow Cytometry of T Cells
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Cultured T cells were washed in PBS/0.1% BSA and re-suspended at 1×106 cells in 50 uL Brilliant Buffer (BD Biosciences) supplemented with 4% rat serum for 15 minutes at 4C. Cells were then incubated for 30 minutes at 4°C in 100 μL of Brilliant Buffer using the following antibody fluorophore conjugates (all from BD Biosciences unless otherwise noted): CD7 BV421, CD4 BV510, CCR4 BV605 (BioLegend), CD8 BV650, CD196 BV786 (BioLegend), CD3 AF488, CD45RA PerCPCy5.5, CD183 PE, CD197 PE-CF594, CD185 PE-Cy7 (BioLegend), and CCR10 APC (R&D Systems). Full details of fluorophore conjugated antibodies can be found in supplemental table 5 . Cells were then washed twice in PBS/0.1% BSA and data acquired on a ZE5 (Yeti) cytometer (BioRad/Propel Labs). Compensation and analyses were performed on FlowJo V10 (TreeStar) using fluorescence minus one (FMO) controls. Statistical analyses were performed on GraphPad Prism 7 using 2-way ANOVA with Bonferroni post-hoc corrections.
4
Multiparametric Flow Cytometry Immunophenotyping
5
Multiparametric Flow Cytometry Immunophenotyping
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Mouse FITC-CD4 and eFluor 605-CD44 [BD-Pharmingen]; KJ1.26 anti-TCR (Caltag Laboratories), Brilliant Violet 570-CD4 and Alexa Fluor 700-CD44 were from [Biolegend], FITC-Klotho,PE-Cy7-CD99, PE-Cy5-VDR, and PE-Cy7-CNR2 were from [Bioss]; APC-CCR10 and APC-ITGA3 were from [R&D Systems]; Fixable viability dye eFluor 780 was from [eBioscience]. Antibodies for human PBMC staining were as follows: from Biolegend, CD99-FITC,CD69-BV421 and CD69-APC , CD45R0-APC and CD45R0-BV421, CD3-AF700, CD4-Pe/Cy7, CD45RA-PE, Itga3 (CD49c)-PE, IL-7R-BV510, CD25-APC, HLA-DR-APC. From BD Biosciences, CCR10-APC and CCR10-PerCP Cy5.5. and viability dye eFluor 780 as above. Relevant mouse anti human isotype antibodies to IgG1k or IgG2ak were utilized for the following fluorophores: BV421, BV510, FITC, PE, and APC for both isotypes (Biolegend).
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