Fresh total cells from spleens were isolated in PBS1×-3% fetal bovine serum (FBS) and stained for 20 min at 4°C with the following monoclonal antibodies at predetermined optimal dilutions: CD121b-BV421, CD19-PeCF594, CD4-V500, CD8a-AF700, Bcl6-APC, CXCR5-Biotin, GL7-e450, CD95 PE, Foxp3-AF488, PD-1-PE (BD Biosciences) or PE-Texas Red (PETR), streptavidin-APC or streptavidin-APC-Cy7 (BD Biosciences), GITR PETR (Miltenyi). CXCR5 staining was performed using biotinylated anti-CXCR5 for 30 min at 20°C followed by APC-or APC-Cy7-labeled streptavidin at 4°C. Intracellular detection of Foxp3 was performed on fixed and permeabilized cells using appropriate buffer (eBioscience), following the manufacturer's recommendations. Stained cells were run on CytoFLEX S cytometer (Beckman -Coulter) and analyzed using FlowJo software (TreeStar Inc.). Dead cells were excluded by forward/side scatter gating.
Bcl6 apc
Manufactured by BD
Sourced in United States
Bcl6-APC is a flow cytometry antibody that targets the Bcl6 protein, a transcriptional repressor involved in the regulation of B-cell differentiation. The APC (Allophycocyanin) fluorescent dye is conjugated to the antibody, enabling its detection and quantification in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Streptavidin apc cy7, by BD (1 mentions)
Foxp3 af488, by BD (1 mentions)
Pd 1 pe, by BD (1 mentions)
Cxcr5 biotin, by BD (1 mentions)
Cd19 pe cf594, by BD (1 mentions)
Cytoflex s cytometer, by Beckman Coulter (1 mentions)
Cd95 pe, by BD (1 mentions)
Pe texas red, by BD (1 mentions)
Streptavidin apc, by BD (1 mentions)
Pd 1 pe cy7, by BD (1 mentions)
Cd138 pe, by BD (1 mentions)
Anti mouse cd4 fitc, by Thermo Fisher Scientific (1 mentions)
Pd 1 pe, by Thermo Fisher Scientific (1 mentions)
Ifn γ pe, by BD (1 mentions)
Foxp3 apc, by BD (1 mentions)
Cxcr5 apc, by Thermo Fisher Scientific (1 mentions)
Cxcr5 pe, by BD (1 mentions)
Cd138 pe, by Thermo Fisher Scientific (1 mentions)
Anti human cd4 fitc, by BD (1 mentions)
Cd25 pe, by BD (1 mentions)
2 protocols using bcl6 apc
1
Multiparametric Flow Cytometric Analysis
2
Multiparametric Immune Cell Profiling
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To examine the expression of surface markers and intracellular molecules, cells were incubated with FcR blocking reagent (Miltenyi, Germany) for 10 min followed by primary antibodies on ice in the dark for 30 min. The antibodies used for surface marker analysis included anti-human CD4-FITC, CXCR5-PE, PD-1-PE-cy7, BCL-6-APC, CD25-PE, CD19-FITC, CD138-PE (BD Pharmingen, USA), and anti-mouse CD4-FITC, CXCR5-APC, PD-1-PE, and CD138-PE (eBioscience, USA). For intracellular staining, cells were cytofixed and cytopermed using the CytofixCytoperm Plus kit (BD Pharmingen, USA) and stained with intracellular antibodies, including anti-human IL-17-APC, IFN-γ-PE, IL-4-PE-cy5, Foxp3-APC (BD Pharmingen, USA), 5-mC and 5-hmC (Abcam, USA) for an additional 30 min on ice in the dark. For some experiments, apoptosis was detected using a FITC Annexin V Apoptosis Detection Kit II (BD Pharmingen, USA). Data were acquired by flow cytometry (BD, Canto II, USA) and analyzed using FlowJo (Tree Star, USA).
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