Approximately 250,000 PBMC were stained with monoclonal antibodies (Becton-Dickinson, San Diego, CA, USA): anti-CD3 [fluorescein isothiocyanate (FITC)], anti-CD8 [peridinin chlorophyll protein (PerCP)] and anti-CD38 [phycoerythrin (PE)]; and anti-CD19 [peridinin chlorophyll protein (PerCP)], anti-CD10 [phycoerythrin (PE)-TexasRed], anti-CD21 [fluorescein isothiocyanate (FITC)], anti-CD27 [phycoerythrin (PE)-Cy7], anti-CD80 [allophycocyanin-H7 (APC-H7)] and anti-CD86 [allophycocyanin (APC)]. Appropriate isotype controls (mouse IgG1-PE and mouse IgG2b-APC) were used to evaluate non-specific staining. All the samples were analyzed by LSR II cytofluorimeter (Becton-Dickinson). A total of 50,000 events was collected in the lymphocyte gate using morphological parameters (Forward and Side scatter). Data were processed with FACSDiva™ Software (Becton-Dickinson) and analysed using Kaluza® Analyzing Software v.1.2 (Beckman Coulter, Fullerton, CA, USA).
Anti cd21
Manufactured by BD
Sourced in United States
Anti-CD21 is a laboratory reagent used in flow cytometry and other immunological applications. It is designed to specifically bind to the CD21 receptor, which is expressed on the surface of certain immune cells. The primary function of Anti-CD21 is to facilitate the identification and analysis of these CD21-positive cells, enabling researchers to study their role in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Anti igd, by BD (4 mentions)
Facsdiva software, by BD (3 mentions)
Anti b220, by BD (3 mentions)
Anti cd3, by BD (2 mentions)
Kaluza analyzing software v1, by Beckman Coulter (2 mentions)
Anti cd8, by BD (2 mentions)
Anti cd19, by BD (2 mentions)
Anti cd27, by BD (2 mentions)
Anti cd138, by BD (2 mentions)
Anti thy1, by BD (2 mentions)
Anti cd43, by BD (2 mentions)
Anti cd38, by BD (1 mentions)
Lsr 2 cytofluorimeter, by BD (1 mentions)
Lsrii flow cytometer, by BD (1 mentions)
Live dead fixable near ir dead cell stain kit, by Thermo Fisher Scientific (1 mentions)
Anti cd4, by BD (1 mentions)
Anti cd19, by Miltenyi Biotec (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Facscalibur, by BD (1 mentions)
Cellquest software, by BD (1 mentions)
7 protocols using anti cd21
1
Comprehensive PBMC Immunophenotyping Assay
2
Comprehensive Immunophenotyping of Cryopreserved PBMC
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Immunophenotyping was performed on cryopreserved PBMC. Cells were thawed, washed, stained for 20 min in the dark with the Live/Dead Fixable Near-IR Dead Cell Stain Kit (Life Technologies, Carlsbad, California, USA) and the following labelled monoclonal antibodies (mAbs): anti-CD3 [fluorescein isothiocyanate (FITC)], anti-CD4 [peridinin chlorophyll protein (PerCP)], anti-CD38 [phycoerythrin (PE)], anti-HLA-DR [allophycocyanin (APC)], anti-CD279 (programmed cell death 1, PD-1) [PE-Cy7], anti-CD57 [PE], anti-CD21 [BV421], anti-CD27 [PE-Cy7], anti-IgD [PE], anti-CD274 (programmed cell death ligand 1, PD-L1) [BV421] (Becton-Dickinson, San Diego, California, USA); anti-CD8 [VioGreen], anti-CD28 [APC], anti-CD19 [VioBright515], anti-CD10 [APC] (Miltenyi Biotec, Auburn, California USA). Cells were then washed and resuspended in phosphate-buffered saline supplemented with 1% paraformaldehyde. All samples were analyzed using LSRII Flow cytometer (Becton-Dickinson). A total of 50000 events were collected in the lymphocyte gate using morphological parameters (forward and side-scatter). Data were processed with FACSDiva Software (Becton-Dickinson) and analyzed using Kaluza Analyzing Software v.1.2 (Beckman Coulter) ( Figure 1 supplementary ).
3
Multiparameter Flow Cytometry of Immune Cells
4
Multiparameter Flow Cytometry Analysis
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Single cell suspensions were prepared and washed in ISCOVES medium. After centrifugation, erythrocytes were lysed in Red Cell Removal Buffer (RCRB; 156 mM NH4Cl, 10 μM EDTA, 1 mM Na2CO3) and FCS was subsequently added (3 (link)). Cells were counted and 106 cells per sample were used for staining. Cells were washed twice in PBS containing 3% FCS and 0.1% NaN3 and were subsequently stained with optimal concentrations of anti-IgM, anti-IgD, anti-B220, anti-CD25, anti-CD21, anti-CD43, anti-Thy 1.2, anti-NK1.1, or anti-CD138 (all from BD Bioscience). Fluorochrome-labeled streptavidin and the apoptosis marker merocyanine (Sigma Aldrich, Munich, Germany) were incubated separately. Samples were subsequently acquired on a FACSCalibur (BD Bioscience) and analyzed using the CellQuest software (BD Bioscience).
5
Multiparametric Flow Cytometry Analysis
6
Comprehensive B Cell Immunophenotyping
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Bone marrow, spleen, mesenteric lymph node (mLN) and Peyer’s Patch (PP) cells were isolated as described previously (26 (link)). In brief, cells were stained with anti-B220 (BioLegend), anti-CD11b (BD Biosciences), anti-CD43 (eBioscience), anti-CD24 (BioLegend), anti-BP1 (eBioscience), anti-IgD (BioLegend), anti-IgM (BioLegend), anti-CD93 (eBioscience) and anti-CD23 (BioLegend) for B cell development staining. For B1 and B2 cell development staining, cells were stained with anti-B220 (BioLegend), anti-CD19 (BioLegend), anti-CD43 (eBioscience), anti-CD23 (BioLegend) and anti-CD5 (BioLegend). For FOB and MZB cell staining, cells were stained with anti-B220 (BioLegend), anti-CD19 (BioLegend), anti-CD21 (BD Biosciences) and anti-CD23 (BioLegend).Cells were stained with anti-B220 (BioLegend), anti-Gl7 (BioLegend), anti-CD95 (Fas) (BD Biosciences), anti-CD86 (BioLegend), and anti-CXCR4(BioLegend) for GC B cell staining and cells were stained with anti-B220 (BioLegend), anti-MHC II (eBioscience) and anti-CD86 (BioLegend) for B cell activation staining.
7
Flow Cytometric Analysis of Class Switch Recombination
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For CSR from IgM to IgA, CH12F3-2A cells were stained with Dylite488-conjugated anti-mouse IgA (Abcam, Cambridge, UK) and APC-conjugated anti-mouse IgM (BD Pharmingen). For CSR from primary B cells, cells were stained with fluorochrome-conjugated anti-IgG1, anti-IgG3, anti-IgE, anti-CD19, anti-B220, anti-CD43, anti-BP1, anti-CD24, anti-CD3, anti-IgD, anti-CD21, and anti-CD23 antibodies (all from BD Pharmingen). Cells were then processed on a BD FACSCanto flow cytometry system and the data were analyzed using FlowJo software (BD Pharmingen).
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