PBMCs were stained with CD4-FITC, CXCR5-PE, CXCR3-APC, CCR6-PE, CD27-FITC, CD38-PE, CD40L-PE, and IFNγ-PE antibodies (BDPharmingenTM, USA) and incubated at 4°C for 30 min in the dark. After the antigen–antibody incubation was completed, the specimen was cleaned 3 times with 2.5 mL of flowing washing solution (Hyclone, USA)/PBS, then resuspended in 500 μL of PBS, and put in storage at 4°C for performing upper flow cytometry analysis. The isotype control antibody was used to adjust the compensation of each channel and set the gate parameters. FlowJo software (Version 7.6.1, Tree Star Inc., USA) was employed to analyze the data obtained by flow cytometry. CD4+CXCR5+CXCR3+CCR6 indicates Th1-like cTfh cells, CD4+CXCR5+CXCR3-CCR6 indicates Th2-like cTfh cells, and CD4+CXCR5+CXCR3-CCR6+ indicates Th17-like cTfh cells.
Ifnγ pe antibodies
Manufactured by BD
Sourced in United States
The IFNγ-PE antibodies are a type of laboratory reagent used for the detection and analysis of interferon-gamma (IFNγ) in various biological samples. These antibodies are conjugated with the fluorescent dye phycoerythrin (PE), which allows for the fluorescent labeling and identification of IFNγ-positive cells through techniques such as flow cytometry.
Automatically generated - may contain errors
2 protocols using ifnγ pe antibodies
1
Multiparameter flow cytometry for cTfh subsets
2
Cytokine Profiling of SARS-CoV-2 RBD Stimulated Splenocytes
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The secretions of IFN-γ, IL-4, and IL-2 in the splenocytes were measured using flow cytometry. The density of splenocytes was adjusted to 2.5 × 106 cells/mL, stimulated with 10 μg/mL of SARS-CoV-2 RBD protein for 20 h, and incubation was done Protein Transport Inhibitor Cocktail for 4 h. The plates were washed three times with a wash buffer. CD3-BV510, CD4-FITC, and CD8-BV785 antibodies were used to stain the splenocytes. The cells were gently mixed with these antibodies and incubated at 4 °C for 30 min in the dark. Then we added Fixation/Permeabilization solution in the dark, and incubation proceeded for 20 min at 4 °C. Furthermore, IL-2-APC, TNF-BV421, and IFN-γ-PE antibodies (BD Biosciences) were used to stain the splenocytes. Finally, the fluorescent signal was detected by using the ACEA system.
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