Control Sirt3+/+ and Sirt3−/− mice were immunized intraperitoneally at 8 to 12 weeks of age with 0.5 ml suspension of 2% sheep red blood cell (SRBC) in PBS (Cocalico Biologicals) to induce GC formation. Mice were sacrificed after 10 days and spleens were isolated from control and SIRT3 knockout mice. The spleen sections derived from Sirt3+/+ and Sirt3−/−mice were stained by hematoxylin and eosin (FI&E) and peanut agglutinin (PNA) using standard procedures. The number of GCs, the total spleen area occupied by GCs and the average area occupied by the GCs were quantified using ImageJ 1,44o (NIH) software. To determine the percentage of GC B-cell population, single-cell suspensions from spleens derived from Sirt3+/+ and Sirt3−/− mice were stained using the following fluorescent-labeled anti-mouse antibodies: FITC conjugated anti-B220, PE conjugated anti-FAS, APC conjugated anti-CD38 from BD biosciences and analyzed by flow-cytometry. To evaluate follicular and marginal zone B-cell populations, splenic B cells were stained with APC conjugated anti-B220, FITC conjugated anti-CD21 and PE conjugated anti-CD38 from BD biosciences and then analyzed by flow-cytometry. DAPI was used for the exclusion of dead cells. The data was analyzed by FlowJo software.
Fitc conjugated anti cd21
Manufactured by BD
The FITC conjugated anti-CD21 is a laboratory reagent used for the detection and analysis of CD21 expression on cells. CD21 is a cell surface receptor that plays a role in B-cell activation and function. The FITC (Fluorescein Isothiocyanate) fluorescent label allows for the visualization and identification of cells expressing CD21 using flow cytometry or other fluorescence-based techniques.
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Lab products found in correlation
Fitc conjugated anti b220, by BD (1 mentions)
Pe conjugated anti fas, by BD (1 mentions)
Apc conjugated anti cd38, by BD (1 mentions)
Pe conjugated anti cd38, by BD (1 mentions)
Apc conjugated anti b220, by BD (1 mentions)
Apc conjugated anti cd19, by BD (1 mentions)
Apc conjugated anti cd11b, by BioLegend (1 mentions)
Pe conjugated anti cd23, by Thermo Fisher Scientific (1 mentions)
Fitc conjugated anti igm, by BD (1 mentions)
Bv605 conjugated streptavidin, by BioLegend (1 mentions)
Ra3 6b2, by BD (1 mentions)
Ra3 6b2, by BioLegend (1 mentions)
Fitc conjugated anti human cd8, by BD (1 mentions)
Pe conjugated anti hla dr, by BD (1 mentions)
Pe conjugated anti cd19, by BD (1 mentions)
Pe conjugated anti cd20, by BD (1 mentions)
Pe cy7 conjugated anti cd38, by BD (1 mentions)
Pe conjugated anti pd 1, by Thermo Fisher Scientific (1 mentions)
Facsdiva, by FlowJo (1 mentions)
Apc conjugated anti human cd8, by BD (1 mentions)
3 protocols using fitc conjugated anti cd21
1
Sirt3 Regulates Germinal Center Formation
2
Multiparameter Flow Cytometry of Immune Cells
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PE-conjugated anti-CD23 (eBioscience-B3B4), FITC-conjugated anti-IgM (Sigma-μ chain specific) or brilliant violet (BV) 421-conjugated anti-IgM (BD pharmingen-R6-60.2), APC-conjugated anti-CD19 (BD bioscience-1D3), FITC-conjugated anti-CD3ε (Immunotools-145-2C11), APC-conjugated anti-CD11b (Biolegend- M1/70) and PE-conjugated anti-Gr-1(Immunotools- RB6-8C5) were used for flow cytometric analysis of blood leukocytes. Biotinylated anti-B220 (eBioscience-RA3-6B2) or Alexa 647-conjugated anti-B220 (BD Pharmingen- RA3-6B2), PE-conjugated anti-IgD (Biolegend-clone 11-26c.2a), FITC-conjugated anti-IgM (Sigma-μ chain specific) or BV 421-conjugated anti-IgM (BD Pharmingen- R6-60.2), FITC- conjugated anti-CD21 (BD Pharmingen- eBio8D9), PE-conjugated anti-CD23 (eBioscience-B3B4), APC-conjugated anti-CD19 (BD bioscience-1D3) or Alexa 700-conjugated anti-CD19 (BD Pharmingen-1D3) were used for flow cytometric analysis of BM or spleen cells. Biotinylated antibodies were detected with APC-conjugated streptavidin (Immunotools) or BV 605-conjugated streptavidin (BioLegend). Rat anti-mouse SIRPα (mAb P84; rat IgG1; a generous gift from Dr.Carl Lagenaur, university of Pittsburgh) was purified and conjugated to Alexa 488 as previously described [18 (link)].
3
Phenotypic Analysis of XABCL-LCL and T-Cells
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Between 2 and 5 × 105 of cells were stained with anti-human antibodies for 20 min at 4°C in the dark with PBS containing 2% FBS. After incubation, cells were washed twice, collected in 200 μl of PBS, and analyzed by flow cytometry.
For the XABCL-LCL phenotype analysis, anti-human antibodies used were PE-conjugated anti-CD19 (BD Pharmingen), PE-conjugated anti-CD20, FITC-conjugated anti-CD21, PE-Cy7-conjugated anti-CD38, APC-conjugated anti-HLA-ABC (BD Pharmingen), PE-conjugated anti-HLA-DR (BD Pharmingen), FITC-conjugated anti-CD80 (ImmunoTools), FITC-conjugated anti-CD86 (ImmunoTools), PE-conjugated anti-PD-1 (eBioscience), and FITC-conjugated anti-PD-L1 (BD Pharmingen). For the T-cell phenotype analysis, the following human antibodies were used: PerCP-conjugated anti-human CD3, PE-conjugated anti-human or FITC-conjugated anti-human CD4, and FITC-conjugated anti-human CD8 or APC-conjugated anti-human CD8 (BD Pharmingen). The flow cytometer used was BD FACS Canto. Analyses were performed by using the FlowJo and FACS Diva software.
For the XABCL-LCL phenotype analysis, anti-human antibodies used were PE-conjugated anti-CD19 (BD Pharmingen), PE-conjugated anti-CD20, FITC-conjugated anti-CD21, PE-Cy7-conjugated anti-CD38, APC-conjugated anti-HLA-ABC (BD Pharmingen), PE-conjugated anti-HLA-DR (BD Pharmingen), FITC-conjugated anti-CD80 (ImmunoTools), FITC-conjugated anti-CD86 (ImmunoTools), PE-conjugated anti-PD-1 (eBioscience), and FITC-conjugated anti-PD-L1 (BD Pharmingen). For the T-cell phenotype analysis, the following human antibodies were used: PerCP-conjugated anti-human CD3, PE-conjugated anti-human or FITC-conjugated anti-human CD4, and FITC-conjugated anti-human CD8 or APC-conjugated anti-human CD8 (BD Pharmingen). The flow cytometer used was BD FACS Canto. Analyses were performed by using the FlowJo and FACS Diva software.
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