Flurochrome-conjugated or biotinylated antibodies against mouse CD4 (RM4-5), CD8 (53-6.7), Foxp3 (FJK-16s), IFN-γ (XMG1.2), IL-4 (BVD6-24G2), NK1.1 (PK136), TCRβ (H57-595), CD45 (30-F11), CD44 (IM7), CD62L (MEL-14), CD11b (M1/70), B220 (RA3-6B2), XCR1 (ZET), NKp46 (29A1.4), CD11c (N418), Ly6c (AL-21), Ly6G (1A8), MHC-II I-A/I-E (M5/114.15.2) were purchased from BD Biosciences, TonBo, eBioscience, Invitrogen and BioLegend. All antibodies were tested with their respective isotype controls. Cell-surface staining was conducted by incubating cells with antibodies for 30 min on ice in the presence of 2.4G2 mAb to block FcγR binding. For Foxp3 staining, a transcription factor-staining kit (Tonbo Biosciences) was used. To assess cytokine production, T cells were stimulated with 50 ng/mL phorbol 12-myristate 13-acetate (Sigma), 1 mM ionomycin (Sigma) in the presence of Golgi-Stop (BD Biosciences) for 4 hr at 37°C as previously described35 . T cells were subsequently stained for cell surface markers before intracellular cytokine staining. All data were acquired using an LSRII flow cytometer (Becton Dickinson) and analyzed with FlowJo software (Tree Star, Inc.).
Mhc 2 1 a 1 e m5 114.15.2
Manufactured by BD
Sourced in United States
The MHC-II I-A/I-E (M5/114.15.2) is a monoclonal antibody that binds to the major histocompatibility complex (MHC) class II I-A and I-E molecules. It is commonly used in flow cytometry and other immunological applications to detect and analyze cells expressing these MHC class II molecules.
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Lab products found in correlation
Lsrii flow cytometer, by BD (1 mentions)
Ionomycin, by Merck Group (1 mentions)
Golgistop, by BD (1 mentions)
Phorbol 12 myristate 13 acetate, by Merck Group (1 mentions)
Rm4 5, by BD (1 mentions)
Bvd6 24g2, by BD (1 mentions)
Transcription factor staining kit, by Cytek Biosciences (1 mentions)
Fjk 16s, by BD (1 mentions)
Cd16 cd32 fc block 2.4g2, by BD (1 mentions)
F4 80, by Thermo Fisher Scientific (1 mentions)
Nkp46 29a1.4, by BioLegend (1 mentions)
Cd4 gk1, by BioLegend (1 mentions)
Cd19 1d3, by BD (1 mentions)
Ly6g 1a8, by BD (1 mentions)
Cd64 x54 5 7, by BioLegend (1 mentions)
Cd11b m1 70, by BD (1 mentions)
Cd45 30 f11, by BD (1 mentions)
Cd11c n418, by BioLegend (1 mentions)
Cd3 17a2, by BD (1 mentions)
Cd90.2 53 2 1, by BD (1 mentions)
3 protocols using mhc 2 1 a 1 e m5 114.15.2
1
Flow Cytometry Analysis of Immune Cells
2
Flow Cytometry Immunophenotyping Panel
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TLR9 (J15A7; BD Biosciences; San Jose, CA), IgG1 (MOPC-21; BD Biosciences; San Jose, CA), CD45 (30-F11; BD Biosciences; San Jose, CA), CD11b (M1/70; BD Biosciences; San Jose, CA), CD11c (N418; Biolegend; San Diego, CA), Siglec F (E50-2440; BD Biosciences; San Jose, CA), MHC II (I-A/I-E) (M5/114.15.2; BD Biosciences; San Jose, CA), CD64 (X54-5/7.1; Biolegend; San Diego, CA), F4/80 (BM8 eBiosciences; SanDiego, CA), LY6G (1A8; BD Biosciences; San Jose, CA), CD3 (17A2, BD Biosciences; San Jose, CA), CD90.2 (53–2.1; BD Biosciences; San Jose, CA), CD4 (GK1.5; Biolegend; San Diego, CA) CD8 (53–6.7; BD Biosciences; San Jose, CA), NKP46 (29A1.4; Biolegend; San Diego, CA), CD19 (1D3; BD Biosciences; San Jose, CA), Fc Block(CD16/CD32) (2.4G2; BD Biosciences; San Jose, CA). The NP antibody used was MA1-7322 from Thermofisher (Waltham, MA) conjugated to FITC.
3
Immunophenotyping of Mouse Immune Cells
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Cells were treated with anti-mouse CD16/CD32 (BioLegend) to block Fc receptors before staining. Staining was performed in PBS solution containing 5% FCS. For all experiments, dead cells were excluded using an Aqua Live/Dead fixable staining reagent (Invitrogen, Waltham, MA, USA). Monoclonal antibodies CD4 (GK1.5), CD5 (53-7.3), CD8 (53-6.7), CD11b (M1/70), CD11c (N418), CD19 (6D5), CD45 (30F11), CD107a (1D4B), Cx3cr1 (SA011F11), Vβ5.1 (MR9-4), Vβ8 (F23.1), Eomes (Dan11Mag), TCRβ (H57-597), and MHC II I-A/I-E (M5/114.15.2) were obtained either from BD, BioLegend, or eBioscience (San Diego, CA, USA). For surface staining, cells were incubated with monoclonal antibodies for 30 min on ice. In the cases of Eomes intracellular staining for CNS cells, fresh isolated cells were used without re-stimulation. Flow cytometric analysis was carried out on using a FACS Canto II (BD) with a FACS Diva software, and data were analyzed using a FlowJo (Tree Star, Waltham, MA, USA) software V10.7.
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