B cells were washed once in cold PBS containing 0.1% BSA (FACS buffer) before blocking with anti-FcγRII/FcγRIII (2.4G2, BD Pharmingen, San Diego, CA). Stainings were performed on ice using conjugated mAbs (eBioscience, San Diego, CA, if not stated otherwise), diluted 1:300 in FACS buffer for surface marker and 1:200 for intracellular cytokine staining followed by incubation for 20 min. After washing with FACS buffer, cells were analysed on a FACS Canto II (BD) using FlowJo software (Tree star, Ashland, OR). The following Abs were used: B220-FITC (#11-0452-86), CD5-PE-Cy7 (#25-0051-81), CD1d-PE (#12-0011-81), CD138-APC (#142506, Biolegend), IgM-FITC (#11-5890-85), IgG1-PE (#12-4015-82), CD4-FITC (#11-0041-82), IL-2-APC (#17-7021-81), IFNγ-APC (#17-7311-82) and IL-10-PerCP (#45-7101-80). Abs against TNF−α−PE (#130-092-245) and IL-17-PE (#130-094-296) were from Miltenyi Biotec. For intracellular staining, the fixation and permeabilization kit (Plus Brefeldin A; eBioscience, Cat. no. 88-8823-88) was used according to manufacturer's recommendation.
Plus brefeldin a
Manufactured by Thermo Fisher Scientific
Plus Brefeldin A is a chemical compound used in cell biology research. It functions by inhibiting the transport of proteins from the endoplasmic reticulum to the Golgi apparatus, leading to the accumulation of proteins within the endoplasmic reticulum.
Automatically generated - may contain errors
Lab products found in correlation
Anti fcγrii fcγriii 2.4g2, by BD (2 mentions)
Il 17 pe, by Miltenyi Biotec (2 mentions)
Facscanto 2, by BD (2 mentions)
Ifn γ apc, by BioLegend (1 mentions)
Cd4 fitc, by BioLegend (1 mentions)
Igg1 pe, by BioLegend (1 mentions)
Il 2 apc, by BioLegend (1 mentions)
Igm fitc, by BioLegend (1 mentions)
Cd138 apc, by BioLegend (1 mentions)
Cd5 pe cy7, by BioLegend (1 mentions)
B220 fitc, by BioLegend (1 mentions)
Cd1d pe, by BioLegend (1 mentions)
T bet pe, by Miltenyi Biotec (1 mentions)
2 protocols using plus brefeldin a
1
B Cell Immunophenotyping and Cytokine Analysis
2
Multiparametric T Cell Phenotyping
Check if the same lab product or an alternative is used in the 5 most similar protocols
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T cells were washed once in cold PBS containing 0.1% BSA (FACS buffer) before blocking with anti-FcγRII/FcγRIII (2.4G2, BD Pharmingen, San Diego, CA). Stainings were performed on ice using conjugated mAbs (eBioscience, San Diego, CA, if not stated otherwise), diluted 1:300 (1:200 for intracellular cytokines) in FACS buffer followed by incubation for 20 min. After washing with FACS buffer, cells were analyzed on a FACS Canto II (BD) and FlowJo software (Tree star, Ashland, OR). The following Abs were used: CD8-FITC (#11-0081-82), IL-2-APC (#17-7021-81), IFNγ-APC (#17-7311-82), CD62L-PE (#12-0621-81), CD44-APC (#17-0441-81), Perforin-APC (#17-9392-80), Granzyme B-PE (#12-8898-80). Abs against TNF-PE (#130-092-245), T-bet-PE (#130-098-653) and IL-17-PE (#130-094-296) were from Miltenyi Biotec. For intracellular staining, the fixation and permeabilization kit (Plus Brefeldin A; eBioscience, Cat. no. 88-8823-88) was used according to manufacturer’s recommendation.
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