Flow cytometry was performed as previously described Fang et al. (2010 (link)). The following antibodies were used: anti-CD3e (clone 145-2C11, Biolegend), anti-CD11b (clone M1/70, Biolegend), anti-CD19 (clone 1D3, BD Biosciences), anti-CD27 (clone LG.3A10, Biolegend), anti-CD29 (clone HMβ1-1, Biolegend San Diego, CA), anti-CD44 (clone IM7, Biolegend), anti-CD45.1 (clone A20, Biolegend), anti-CD45.2 (clone 104, Biolegend), anti-CD49a (clone HMα1, Biolegend), anti-CD49b (clone DX5, Biolegend), anti-CD105 (clone MJ7/18, Biolegend), anti-CD106 (clone 429, Biolegend), anti-BrDU (clone PRB-1, eBioscience), anti-CXCR3 (clone CXCR3-173, Biolegend), anti-Eomes (clone Dan11mag, eBioscience), anti-KLRG1 (clone 2F1/KLRG1, Biolegend), anti-Ly-6A/E (Sca-1) (clone E13-161.7, Biolegend), anti-Ly49A (clone A1/Ly49A, Biolegend), anti-Ly49C/I (clone 5E6, Biolegend), anti-Ly49D (clone 4E5, Biolegend), anti-Ly49G2 (clone LGL-1, eBioscience), anti-Ly49H (clone 3D10, Biolegend), anti-NK1.1 (clone PK136, Biolegend), NKG2A/C/E (clone 20d5, Biolegend), anti-Tbet (clone 4B10, Biolegend), and anti-TRAIL (clone N2B2, Biolegend). At least 500 000 cells were analyzed by flow cytometry at the Fox Chase Cell Sorting Facility using an LSR II system (BD Biosciences).
Anti cd19 clone 1d3
Manufactured by BD
Sourced in United States
Anti-CD19 (clone 1D3) is a monoclonal antibody that binds to the CD19 antigen expressed on the surface of B cells. It is commonly used in flow cytometry and other immunological applications for the identification and analysis of B cells.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd3e clone 145 2c11, by BioLegend (2 mentions)
Anti ly6g clone 1a8, by BioLegend (2 mentions)
Anti cd11b clone m1 70, by BD (2 mentions)
Anti ly6c clone hk1, by BioLegend (2 mentions)
Anti cd49b clone dx5, by BioLegend (1 mentions)
Lsrii system, by BD (1 mentions)
Anti nk1.1 clone pk136, by BioLegend (1 mentions)
Anti cd27 clone lg 3a10, by BioLegend (1 mentions)
Anti cd44 clone im7, by BioLegend (1 mentions)
Anti tbet clone 4b10, by BioLegend (1 mentions)
Anti cd45.1 clone a20, by BioLegend (1 mentions)
Anti cd45.2 clone 104, by BioLegend (1 mentions)
Anti klrg1 clone 2f1 klrg1, by BioLegend (1 mentions)
Anti cxcr3 clone cxcr3 173, by BioLegend (1 mentions)
Anti cd11b clone m1 70, by BioLegend (1 mentions)
Anti cd90 2 clone 53 2, by BD (1 mentions)
Macs neural tissue dissociation kit, by Miltenyi Biotec (1 mentions)
Facsverse, by BD (1 mentions)
Fixable viability stain 510, by BD (1 mentions)
Anti cd3 clone 145 2c11, by BD (1 mentions)
5 protocols using anti cd19 clone 1d3
1
Comprehensive Immune Cell Profiling by Flow Cytometry
2
Immunophenotyping of Myeloid Cells
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Blood was incubated with red blood cell lysis buffer for 4 minutes and washed with FACS buffer; this was repeated twice more. Single-cell suspensions were stained with the following monoclonal antibodies: APC anti-CD115 (clone AFS98, 1:200; eBiosciences), PerCP-Cy5.5 anti-CD11b (clone M1/70, 1:200; BioLegend), PE-Cy7 anti-Ly6C (clone Al-21, 1:200; BioLegend), eFluor 450 anti-Ly6G (clone 1A8, 1:100; eBioscience), anti-CD19 (clone 1D3, 1:400; BD Pharmingen) and anti-CD90.2 (clone 53-2.1, 1:400; BD Pharmingen); 10 μL DAPI was added for viability staining. CD11b+ cells were identified as CD11b+ and Lin−. Neutrophils were identified as CD11bhigh, Lin−/low, CD115low, and Ly6Ghigh. Monocytes were identified as CD11bhigh, Lin−/low, CD115high, and Ly6Chigh/low.
3
Immune Cell Profiling of Post-Operative Mice
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Single-cell suspensions were prepared from the blood and brain tissues of control or surgery group mice at postoperative day 2. Brain tissues were dissociated by MACS Neural Tissue Dissociation Kit (Miltenyi Biotec GmbH, Teterow, Germany) according to the protocol of the manufacturer. The cells were incubated with Fixable Viability Stain 510 (BD PharMingen, San Diego, CA, United States) for 15 min at room temperature protected from light. After that, these cells were stained with fluorochrome-conjugated antibodies. The following mouse antibodies were used in this study: anti-CD45 (clone 104, #560694, BD Biosciences, San Diego, United States), anti-CD11b (clone M1/70, #561098, BD Biosciences, San Diego, United States), anti-Ly6G (clone 1A8, #127614, Biolegend, United States), anti-Ly6C (clone HK1.4, #128007, Biolegend, United States), anti-F4/80 (clone T45-2342, #565411, BD Biosciences, San Diego, United States), anti-CD3 (clone 145-2C11, #551163, BD Biosciences, San Diego, United States), anti-CD19 (clone 1D3, #561738, BD Biosciences, San Diego, United States), anti-NK1.1 (clone PK136, #45-5941-80, Invitrogen, Carlsbad, United States), and anti-CD11c (clone HL3, #561022, BD Biosciences, San Diego, United States). All acquisitions were performed using BD Biosciences FACSVerse™ and analyzed using FlowJo 10.4 software.
4
Detailed FACS Sorting and Analysis
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For FACS sorting and analysis, we used previously described staining protocols (Mossadegh-Keller et al., 2013 (link)), published stem and progenitor cell definitions (Bryder et al., 2006 (link)), FACSCanto, LSRII, and FACSAriaIII equipment, and DIVA software (BD), analyzing only populations with at least 200 events. The following antibodies were used for staining cells: anti-CD117 (clone 2B8; BD), anti–Sca-1 (clone D7; BioLegend), anti-CD34 (clone RAM34; BD), anti-CD16/32 (clone 2.4G2; BD), anti-CD11b (clone M1/70; BD), anti-CD19 (clone 1D3; BD), anti-CD3e (clone 145-2C11; BioLegend), anti-Ly6G (clone 1A8; BioLegend), anti-Ly6C (clone HK1.4; BioLegend), anti-CD115 (clone AFS98; eBioscience), anti-CD45.2 (clone 104; BD), anti-CD45.1 (clone A20; BD), anti-B220 (clone RA3-6B2; eBioscience), anti-Ter119 (clone TER-119; eBioscience), and anti-CD71 (clone R17217; eBioscience). LIVE/DEAD Fixable Violet Dead cell dye (Invitrogen) was used as a viability marker. Information describing gating strategies is shown in Figs. S1 and S2.
5
Comprehensive PBMC Immunophenotyping Panel
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After thawing of PBMCs, cells were washed in dPBS supplemented with 0.5% BSA. The cells were stained with anti-CD19 (clone 1D3, 557398, BD biosciences) and anti-CD56 (Clone CMSSB, 46-0567-42), anti-CD16 (clone eBioB16, 17-0168-42), anti-CD4 (clone OKT4, 56-0048-41), anti-CD8 (clone SK1, 48-0087-42; clone RPA-T8, 56-0088-41) and anti-CD3 (clone OKT3, 86-0037-42) (all eBioscience); anti-NKG2D (clone 1D11, 320806), anti-CD2 (clone TS1/8, 309214), anti-CD3 PE Cy7 (300420) (all Biolegend); anti-CD56 PerCP e71 (46-0566-41) (Thermo Fisher Scientific). FVD 780 APC-Cy7 (Thermo Fisher Scientific) was used for live/dead staining.
Data were collected on a BD LSR Fortessa and analysed using FlowJo V.10.5 (Tree Star Inc.).
Data were collected on a BD LSR Fortessa and analysed using FlowJo V.10.5 (Tree Star Inc.).
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