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2 protocols using percpfl710

1

Multiparametric Flow Cytometry Analysis

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The following flurochrome-conjugated antibodies were used for surface staining: anti-human CD3 (Brilliant Violet 650, 1:100, OKT3, BioLegend, San Diego, CA), CD4 (PeCy7, 1:100, SK3, BioLegend), CD8 (APC-Cy7, 1:100, SK1, BD Biosciences), CD19 (PE Texas Red, 1:100, SJ25-C1, Invitrogen or APC, HIB19, Biolegend), CD25 (FITC, 1:50, 2A3, BD Biosciences, San Jose, CA), CD31 (APC, 1:100, WM59, eBioscience, San Diego, CA), CD45RA (Brilliant Violet 605, 1:100, HI100, BioLegend), CD45RO (PerCpFl710, 1:100, UCHL1, eBioscience), CD69 (Brilliant Violet 421, 1:100, FN50, BioLegend or BUV395, 1:100, FN50, BD Biosciences), CD103, (Alexa Fluor 647, 1:100, Ber-ACT8, BioLegend), CD127 (BV711, 1:100, A019D5, BioLegend), CCR7 (Alexa Fluor 488, 1:100, TG8, BioLegend). For intracellular staining, surface stained cells were resuspended and incubated in fixation buffer (eBioscience), washed, resuspended in 0.1mL permeabilization buffer (eBioscience) and stained with anti-Foxp3 antibodies (PE, 1:20, 236A/E7, eBioscience) and Ki67 (α700, 1:100, Ki-67, BioLegend) for 30 min at room temperature and washed twice with permeabilization buffer. Stained cells were acquired on a 6-laser LSRII analytical flow cytometer (BD Biosciences) in the CCTI flow cytometry core and analyzed using FlowJo software (Treestar, Ashland, OR).
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2

Multiparametric Flow Cytometry Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
The following flurochrome-conjugated antibodies were used for surface staining: anti-human CD3 (Brilliant Violet 650, 1:100, OKT3, BioLegend, San Diego, CA), CD4 (PeCy7, 1:100, SK3, BioLegend), CD8 (APC-Cy7, 1:100, SK1, BD Biosciences), CD19 (PE Texas Red, 1:100, SJ25-C1, Invitrogen or APC, HIB19, Biolegend), CD25 (FITC, 1:50, 2A3, BD Biosciences, San Jose, CA), CD31 (APC, 1:100, WM59, eBioscience, San Diego, CA), CD45RA (Brilliant Violet 605, 1:100, HI100, BioLegend), CD45RO (PerCpFl710, 1:100, UCHL1, eBioscience), CD69 (Brilliant Violet 421, 1:100, FN50, BioLegend or BUV395, 1:100, FN50, BD Biosciences), CD103, (Alexa Fluor 647, 1:100, Ber-ACT8, BioLegend), CD127 (BV711, 1:100, A019D5, BioLegend), CCR7 (Alexa Fluor 488, 1:100, TG8, BioLegend). For intracellular staining, surface stained cells were resuspended and incubated in fixation buffer (eBioscience), washed, resuspended in 0.1mL permeabilization buffer (eBioscience) and stained with anti-Foxp3 antibodies (PE, 1:20, 236A/E7, eBioscience) and Ki67 (α700, 1:100, Ki-67, BioLegend) for 30 min at room temperature and washed twice with permeabilization buffer. Stained cells were acquired on a 6-laser LSRII analytical flow cytometer (BD Biosciences) in the CCTI flow cytometry core and analyzed using FlowJo software (Treestar, Ashland, OR).
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