Splenocytes, tumor single-cell suspensions, or peripheral blood cells were washed with flow buffer (PBS with 1% of FBS and 2 mmol/L of EDTA), then incubated with an Fc-blocking antibody (anti-mouse CD16/ CD32 mAb 2.4G2; BD Biosciences) and stained with fluorescence-conjugated antibodies against surface markers. Cells were then fixed in Fix/Perm buffer (eBioscience) and stained with antibodies against intracellular proteins. The following fluorochrome-labeled antibodies were used: Gr1 (clone RB6-8C5), CD11b (clone M1/70), Ly6G (clone 1A8), Ly6C (clone AL-21), F4/80 (clone BM8), CD206 (clone C068C2), Arg 1 (polyclonal antibody, R & D systems®, Cat: IC5868A), iNOS (clone CXNFT), CD4 (clone RM4-5), CD8a (clone 53-6.7), TCR-Vβ3 (clone KJ25), and TCR-Vβ8 (clone F23.1) were purchased from BD Biosciences (San Jose, CA), eBioscience (San Diego, CA), BioLegend (San Diego, CA), and R & D systems. Cells stained with specific antibodies, as well as isotype-control stained cells, were assayed on a FACScalibur, a FACSCantoII, or a LSR II flow cytometer (BD Biosciences). Data analysis was performed using the FCS Express (De Novo Software) or FACS Diva software (BD Biosciences).
Mab 2.4g2
Manufactured by BD
MAb 2.4G2 is a mouse monoclonal antibody that binds to the Fc gamma receptor II (FcγRII). It is commonly used as a research tool in flow cytometry and immunohistochemistry applications.
Automatically generated - may contain errors
Lab products found in correlation
Facscanto 2, by BD (1 mentions)
Lsrii flow cytometer, by BD (1 mentions)
Facscalibur, by BD (1 mentions)
Facsdiva software, by BD (1 mentions)
Fix perm buffer, by Thermo Fisher Scientific (1 mentions)
Fcs express, by De Novo Software (1 mentions)
Facsverse flow cytometer, by BD (1 mentions)
Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (1 mentions)
Anti human cd3, by BD (1 mentions)
Anti mouse cd3, by BD (1 mentions)
Anti cd4, by Thermo Fisher Scientific (1 mentions)
Anti cd11b, by Thermo Fisher Scientific (1 mentions)
Anti cd206, by BioLegend (1 mentions)
Anti cd4, by BD (1 mentions)
Anti gata3, by Thermo Fisher Scientific (1 mentions)
Anti cd206, by BD (1 mentions)
2 protocols using mab 2.4g2
1
Multiparametric Flow Cytometry for Immune Cell Profiling
2
Quantification of Immune Cell Populations
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Cell suspensions were prepared in PBS containing 2% FBS. Cells (1 × 106) were blocked with mouse Fc blocking solution (anti-mouse CD16/CD32 mAb 2.4G2, BD Bioscience) and stained for 30 minutes at 4 °C with the appropriate antibodies. The antibodies used included: human and mouse anti-Foxp3 (1:50, 150D/E4) and anti-Gata3 (1:50, TWAJ) (eBioscience; San Diego, CA, USA); mouse anti-CD3 (1:50, 145-2C11), anti-CD4 (1:50, GK1.5) (BD Bioscience; F4/80), anti-BM8 (eBioscience), and anti-CD206 (1:50, C068C2) (BioLegend); and human anti-CD3 (1:50, SK7), anti-CD206 (1:50, 19.2) (BD Bioscience), anti-CD4 (1:50, OKT4), and anti-CD11b (1:50, ICRF4) (eBioscience). For intracellular staining, a Foxp3/Transcription factor staining buffer set was purchased from eBioscience. Data were acquired using a FACSVerse flow cytometer (BD Bioscience) and analysed using FlowJo software (Tree Star, San Carlos, CA, USA).
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