Aliquots of 5 x 105 cells were washed in FACS buffer (PBS, 0.1% BSA) and stained using the following panel of monoclonal antibodies (mAbs) to murine cell surface molecules (all from BD Biosciences): PerCP-Cy5.5-conjugated anti-CD11b, PE-conjugated anti-Ly6G, FITC-conjugated anti-Ly6C, allophycocyanin-conjugated anti-CD45. Cells were washed in FACS buffer and data were collected using a two-laser equipped FACSCalibur apparatus and the CellQuest analysis software (Becton Dickinson, Mountain View, CA).
Percp cy5.5 conjugated anti cd11b
Manufactured by BD
Sourced in United States
PerCP-Cy5.5-conjugated anti-CD11b is a fluorochrome-conjugated monoclonal antibody that binds to the CD11b cell surface antigen. CD11b is expressed on the surface of various immune cells, including monocytes, macrophages, and neutrophils. The PerCP-Cy5.5 fluorochrome allows for the detection and analysis of CD11b-positive cells using flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Fitc conjugated anti ly6c, by BD (2 mentions)
Pe conjugated anti ly6g, by BD (2 mentions)
Cellquest analysis software, by BD (1 mentions)
Facscalibur apparatus, by BD (1 mentions)
Cflow plus software, by BD (1 mentions)
Accuri c6 flow cytometer, by BD (1 mentions)
Fc block, by BD (1 mentions)
Facscalibur analyser, by BD (1 mentions)
Cd16 32, by BioLegend (1 mentions)
5 protocols using percp cy5.5 conjugated anti cd11b
1
Murine Immune Cell Phenotyping by Flow Cytometry
2
Murine Immune Cell Phenotyping
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Aliquots of 5 × 105 cells were washed in FACS buffer (PBS, 0.1% BSA), incubated with a fixable viability dye and anti-Fc receptor (αCD16/32), and stained using the following panel of monoclonal antibodies to murine cell surface molecules (all from BD Biosciences): PerCP-Cy5.5-conjugated anti-CD11b, PE-conjugated anti-Ly6G, FITC-conjugated anti-Ly6C, and Violet1-conjugated anti-CD45. Cells were washed in FACS buffer and analyzed by Dako Cyan Flow cytometry.
3
Multicolor Flow Cytometry for Immune Cell Profiling
4
Phagocytosis Assay of MC38 Cells
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For the phagocytosis assay, MC38 cells were treated with A3373 in DMEM containing 1% FBS for 48 h and were labeled with 2 μM carboxyfluorescein succinimidyl ester (CFSE). A total of 5 × 105 THP1 cells were incubated with equal numbers of CFSE-labeled cells in DMEM without FBS for 2 h in a 24-well plate. The cells were washed with PBS and incubated with human BD Fc block (BD Biosciences) to block nonspecific detection. Furthermore, the cells were incubated with PerCP-Cy5.5-conjugated anti-CD11b (BD Biosciences) for 20 min at 4 °C and were analyzed by flow cytometry.
5
Phenotypic Characterization of Blood Immune Cells
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Peripheral blood was collected in anticoagulated tubes, and mononuclear cells were isolated for further analysis. These cell samples were resuspended in RPMI 1640 and incubated in 5% CO 2 at 37 ℃ for 3 hours. All samples were detected on a FACS calibur analyser (Becton-Dickinson, Mountain View, CA).
Then, these cells were incubated with an Fcγ receptor-blocking mAb (CD16/32; BioLegend, San Diego, CA, USA) for 15 minutes at 4 °C. Subsequently, for surface antigen detection, the cells were shielded from light and labeled with with PE-conjugated anti-CD163 (1:100), PE-Cyanine7-conjugated anti-CD206 (Invitrogen, Calif., 1: 100), Alexa Fluor 488-conjugated anti-iNOS (Invitrogen, Calif., USA, 1: 200), PE/Cyanine7-conjugated anti-CD284 (Biolegend, Calif., USA, 1: 100), Alexa Fluor® 647-conjugated anti-F4/80 (BD Biosciences, Franklin Lakes, USA, 1: 100), PE-conjugated anti TIM-1 (BD Biosciences, Franklin Lakes, USA, 1: 100) and PerCP-Cy™5.5-conjugated anti-CD11b (BD Biosciences, Franklin Lakes, NJ, USA, 1: 100) antibodies. All data were analyzed using FlowJo 7.6.1 software (TreeStar, Ashland, OR, USA), and the inactive cells were excluded by gating.
Then, these cells were incubated with an Fcγ receptor-blocking mAb (CD16/32; BioLegend, San Diego, CA, USA) for 15 minutes at 4 °C. Subsequently, for surface antigen detection, the cells were shielded from light and labeled with with PE-conjugated anti-CD163 (1:100), PE-Cyanine7-conjugated anti-CD206 (Invitrogen, Calif., 1: 100), Alexa Fluor 488-conjugated anti-iNOS (Invitrogen, Calif., USA, 1: 200), PE/Cyanine7-conjugated anti-CD284 (Biolegend, Calif., USA, 1: 100), Alexa Fluor® 647-conjugated anti-F4/80 (BD Biosciences, Franklin Lakes, USA, 1: 100), PE-conjugated anti TIM-1 (BD Biosciences, Franklin Lakes, USA, 1: 100) and PerCP-Cy™5.5-conjugated anti-CD11b (BD Biosciences, Franklin Lakes, NJ, USA, 1: 100) antibodies. All data were analyzed using FlowJo 7.6.1 software (TreeStar, Ashland, OR, USA), and the inactive cells were excluded by gating.
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