Experiment Overview
| Repository ID: | FR-FCM-ZZ2M | Experiment name: | Sequential Staining Improves Detection of CCR2 and CX3CR1 on Monocytes When Simultaneously Evaluating CCR5 by Multicolor Flow Cytometry | MIFlowCyt score: | 85.55% |
| Primary researcher: | Emilie Jalbert | PI/manager: | Jason Barbour | Uploaded by: | Emilie Jalbert |
| Experiment dates: | 2011-12-01 - 2012-04-30 | Dataset uploaded: | Aug 2012 | Last updated: | Jan 2013 |
| Keywords: | [multicolor flow cytometry] [chemokine receptor] [antibody staining] [monocyte] | Manuscripts: | [23426986] |
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| Organizations: |
University of Hawaii, Tropical Medicine, Honolulu, Hawaii (USA)
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| Purpose: | Using multicolor flow cytometry, we sought to evaluate and optimize the simultaneous detection of cell surface expression levels of CCR2, CX3CR1 and CCR5 in primary human monocytes using a single antibody panel. | ||||
| Conclusion: | We observed that staining for CCR2 alone or for CX3CR1 alone showed greater expression levels than when the cells were stained with the full panel of antibodies. Fluorescent-minus-one (FMO) controls revealed that ligation of the CCR5 monoclonal antibody to the receptor interfered with detection of CX3CR1 and CCR2. Sequential addition of the antibodies during the staining procedure was sufficient to restore the detection levels, suggesting close proximity and possible functional interactions between CCR2/CCR5 and CX3CR1/CCR5 in monocytes. | ||||
| Comments: | None | ||||
| Funding: | Not disclosed | ||||
| Quality control: | Fluorescence-Minus-One (FMO) and single-stained samples. | ||||
Experiment variables
| Sample Type | |
|---|---|
| · Single Stain | 20111201 Single Stain_CD14 Q605.fcs · 20111201 Single Stain_CD16 Ax700.fcs · 20111201 Single Stain_CD163 PE.fcs · 20111201 Single Stain_CD192 Ax647.fcs · 20111201 Single Stain_CD195 V450.fcs · 20111201 Single Stain_CD3 V500.fcs · 20111201 Single Stain_CD36 FITC.fcs · 20111201 Single Stain_CD56-19-20 PE-Cy7.fcs · 20111201 Single Stain_CX3CR1 PerCP-eF710.fcs · 20111201 Single Stain_HLA-DR APC-H7.fcs · 20111201 Single Stain_Unstained.fcs · 20111201 Single Stain_YARD.fcs |
| · FMO | 20111201 All CCR_D2_D02.fcs · 20111201 All CCR_D3_D03.fcs · 20111201 All CCR_D4_D04.fcs · 20111201 CX-R2-R5_B2_B02.fcs · 20111201 CX-R2-R5_B3_B03.fcs · 20111201 CX-R2-R5_B4_B04.fcs · 20111201 R2-CX-R5_A2_A02.fcs · 20111201 R2-CX-R5_A3_A03.fcs · 20111201 R2-CX-R5_A4_A04.fcs · 20111201 R5-R2-CX_C2_C02.fcs · 20111201 R5-R2-CX_C3_C03.fcs · 20111201 R5-R2-CX_C4_C04.fcs · 20120404 FMO_D11_D11.fcs · 20120404 FMO_D7_D07.fcs · 20120404 FMO_D8_D08.fcs · 20120404 FMO_D9_D09.fcs · 20120411 FMO_FMO CD16_D09.fcs · 20120411 FMO_FMO CD192_D05.fcs · 20120411 FMO_FMO CD195_D07.fcs · 20120411 FMO_FMO CX3CR1_D06.fcs |
| · All Stain | 20111201 All CCR_D1_D01.fcs · 20111201 CX-R2-R5_B1_B01.fcs · 20111201 R2-CX-R5_A1_A01.fcs · 20111201 R5-R2-CX_C1_C01.fcs · 20120404 All Stain_D4_D04.fcs · 20120404 All Stain_D5_D05.fcs · 20120404 All Stain_D6_D06.fcs · 20120411 All Stain_D3_D03.fcs · 20120411 All Stain_D4_D04.fcs |
| · Comps | 20120404 Comps Beads_APC-H7.fcs · 20120404 Comps Beads_Ax647.fcs · 20120404 Comps Beads_Ax700.fcs · 20120404 Comps Beads_FITC.fcs · 20120404 Comps Beads_PE-Cy7.fcs · 20120404 Comps Beads_PerCP-eF710.fcs · 20120404 Comps Beads_Q605.fcs · 20120404 Comps Beads_Unstained.fcs · 20120404 Comps Beads_V450.fcs · 20120404 Comps Beads_V500.fcs · 20120404 Controls_Rainbow Beads 1 peak.fcs · 20120404 Controls_Rainbow Beads 8 peak.fcs · 20120411 Comps Beads_APC-H7.fcs · 20120411 Comps Beads_Ax647.fcs · 20120411 Comps Beads_Ax700.fcs · 20120411 Comps Beads_FITC.fcs · 20120411 Comps Beads_PE-Cy7.fcs · 20120411 Comps Beads_PerCP-eF710.fcs · 20120411 Comps Beads_Q605.fcs · 20120411 Comps Beads_Unstained.fcs · 20120411 Comps Beads_V450.fcs · 20120411 Comps Beads_V500.fcs · 20120411 Controls_Rainbow Beads 1 peak.fcs · 20120411 Controls_Rainbow Beads 8 peak.fcs |
