Spleen and thymus of six-week-old NOD.Foxp3-GFP/cre (Stock No: 008694, Jackson Laboratory) were processed using frosted glass slides and passed through a 40-micron filter to create single cell suspensions. Red blood cells (RBC) were lysed with ammonium-chloride-potassium buffer prior to staining for flow cytometry. Samples were stained with Fixable Live/Dead Near IR (Invitrogen), washed once with stain buffer (PBS + 2% FBS + 0.05% NaN3), and Fc receptors were blocked with anti-CD16/32 for 5 minutes at 4°C (Clone 2.4G2, BD Biosciences). Samples were stained for 30 minutes at 4°C with the following anti-mouse antibodies: CD3e-Brilliant Violet (BV) 605 (145-2C11, BioLegend), CD4-PerCP/Cy5.5 (RM4-5, Thermo Fisher Scientific), CD8a-BV711 (53-6.7, BioLegend), CD44-PE-Cy7 (IM7, BioLegend), CD62L-APC (MEL-14, BioLegend), and CD221-PE (3B7, Santa Cruz Biotechnology). Samples were washed once with stain buffer prior to data acquisition on an LSRFortessa (BD Biosciences) and analysis with FlowJo (v10.6.1; Tree Star).
Cd62l apc mel 14
Manufactured by BioLegend
CD62L-APC (MEL-14) is a fluorescent-conjugated antibody that binds to the CD62L (L-selectin) cell surface marker. CD62L is expressed on various leukocyte subsets, including naive T cells, and plays a role in lymphocyte homing and migration.
Automatically generated - may contain errors
Lab products found in correlation
Foxp3 gfp cre, by Jackson ImmunoResearch (1 mentions)
Cd44 pe cy7 im7, by BioLegend (1 mentions)
Cd221 pe 3b7, by Santa Cruz Biotechnology (1 mentions)
Lsrfortessa, by BD (1 mentions)
Anti cd16 32, by BD (1 mentions)
Clone 2.4g2, by BD (1 mentions)
Cd8α pacific blue, by BioLegend (1 mentions)
Eomes pe, by Thermo Fisher Scientific (1 mentions)
Ifn γ percp cy5, by BioLegend (1 mentions)
Tcrβ apc e780 h57 597, by Thermo Fisher Scientific (1 mentions)
Live dead aqua, by Thermo Fisher Scientific (1 mentions)
Goat anti rabbit igg af488, by Thermo Fisher Scientific (1 mentions)
Pe texas red, by Thermo Fisher Scientific (1 mentions)
2 protocols using cd62l apc mel 14
1
Murine Splenic and Thymic T-cell Profiling
2
Multiparameter Flow Cytometry Analysis
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The following antibodies were used (from BD Biosciences unless otherwise noted): CD8α-Pacific Blue or AlexaFluor (AF)700 (53-6.7, Biolegend); CD4 fluoroscein isothiocyanate (FITC) (GK1.5, eBiosciences), allophycocyanin (APC) (RM4-5), phycoerythrin (PE)-Cy5 or PE-Cy7 (RM4-5, Biolegend), or PE-TexasRed (RM4-5, Invitrogen), TCRβ APC-e780 (H57-597, eBiosciences), IL4Ra PE (mIL4R-M1), CD62L APC (MEL-14),, CD44 AF700 (IM7, Biolegend), CD122-biotin (TM-1) followed by streptavidin-PE Texas Red, Eomes-PE or AF647 (Dan11mag, eBiosciences), IFNγ-PerCPCy5.5 (XMG1.2, Biolegend), APC or PE, pSTAT6 PE (J1-773.58.11), and pAkt T308 PE (J1-223.371). For pS6 staining, cells were stained with a rabbit anti-phospoS6 monoclonal (2F9; Cell Signaling) and then stained with AF488 goat anti-rabbit IgG (Invitrogen). Live/Dead Aqua (Invitrogen) was used to exclude dead cells.
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