Single cell suspensions were prepared as previously described 15 (link), and contaminating erythrocytes were lysed using ammonium chloride buffer where appropriate. Cell suspensions were stained with optimal dilutions of antibodies directly conjugated to either Biotin, FITC, PE, PE-Texas Red, PE-Cy5.5, PerCP-Cy5.5, PerCP-eFluor710, PE-Cy7, Pacific Blue, eFluor 450, allophycocyanin, Alexa Fluor 647, or allophycocyanin-eFluor 780. Biotinylated antibodies were visualized using streptavidin PE-Texas Red (BD Bioscience). Lineage positive cells were gated out of whole BM suspensions using mouse hematopoietic lineage eFluor450 cocktail (eBioscience).The following specific antibodies were used to detect cell surface antigens: CD117/c-Kit (2B8), Sca-1/Ly6A/E (D7), CD127/IL-7Rα (A7R34), CD34 (RAM34), CD135/Flt3 (A2F10), CD25 (PC61.5), AA4.1 (AA4.1), CD27 (LG.7F9), CD150 (mShad150), CD48 (HM48-1), CD244.2 (eBio244F4), CD45.1 (A20), CD45.2 (104), GR-1/Ly-6G (RB6-8C5), CD11b (M1/70), CD11c (N418), CD45R/B220 (RA3-6B2), CD19 (eBio1D3), CD3 (145-2C11), CD4 (GK1.5), and CD8 (53-6.7). All antibodies were obtained from eBioscience. The CD1d PBS-57 tetramer was obtained from the NIH Tetramer Core Facility (Emory University, Atlanta, USA).
Cd3 145 2c11 cd4 gk1.5
Manufactured by Thermo Fisher Scientific
The CD3 (145-2C11) CD4 (GK1.5) is a set of antibodies used for the detection and analysis of mouse T cell subsets. The CD3 antibody binds to the CD3 complex, which is expressed on all mature T cells. The CD4 antibody binds to the CD4 co-receptor, which is expressed on a subset of T cells. These antibodies are commonly used in flow cytometry applications to identify and characterize T cell populations.
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Lab products found in correlation
Cd8 53 6, by Thermo Fisher Scientific (3 mentions)
Foxp3 fix perm kit, by Thermo Fisher Scientific (2 mentions)
Anti mouse ifn γ ab xmg1.2, by BioLegend (2 mentions)
Anti foxp3 mab fjk 16s, by Thermo Fisher Scientific (2 mentions)
Lsrfortessa flow cytometer, by BD (2 mentions)
Streptavidin pe texas red, by BD (1 mentions)
Efluor450 cocktail, by Thermo Fisher Scientific (1 mentions)
3 protocols using cd3 145 2c11 cd4 gk1.5
1
Immunophenotyping of Murine Hematopoietic Cells
2
Multicolor Flow Cytometry Analysis
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Hepatic non-parenchymal cells (NPC) and spleen cells were treated with FcγR-blocking rat anti-mouse CD16/32 mAb (2.4G2) to prevent non-specific antibody (Ab) binding. They were then incubated for 30 min with fluorescein isothiocyanate (FITC)-, phycoerythrin (PE)-, APC-, PE-cyanin (Cy)5-, PE-Cy7-or pacific blue-conjugated monoclonal Abs (mAbs) to detect surface expression of CD3 (145-2C11) CD4 (GK1.5) or CD8 (53–6.7) (all eBioscience, San Diego, CA). For intracellular cytokine staining, cells were fixed with 4% paraformaldehyde and permeabilized using 0.1% saponin, then stained with anti-mouse IFN-γ Ab (XMG1.2) (BioLegend). For forkhead box p3 (Foxp3) staining, cells were fixed and permeabilized using Foxp3 Fix Perm kit (eBioscience) and stained with anti-Foxp3 mAb (FJK-16s) (eBioscience). Appropriate Ig isotype controls were obtained from BD Pharmingen (San Diego, CA). Flow analysis was performed using an LSR Fortessa flow cytometer (BD Biosciences) and results expressed as percent positive cells and mean fluorescence intensity (MFI).
3
Multicolor Flow Cytometry Analysis
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