Peripheral blood mononuclear cells (PBMCs) were isolated from AML patients and HIs and then incubated with the following antibodies: CD45-V450 (clone H130), CD3-Alexa Fluor® 700 (clone SP34-2), TCR γδ-PE-Cy7 (clone B1), Vδ1-FITC (clone TS8.2), Vδ2-PerCP (clone B6), PD-1-PE (clone EH12.2H7), Foxp3-Alexa Fluor® 647 (clone 150D), PE isotype control (clone MOPC-21), and Alexa Fluor® 647 isotype control (clone 259D/C7) (BioLegend, SanDiego, USA; BD Biosciences, San Jose, USA; Abcam Cambridge, UK). First, cells obtained from fresh PB samples were stained with surface markers including CD45, CD3, TCR γδ, Vδ1, Vδ2, and PD-1 at 4°C in the dark for 30 minutes. For Foxp3 expression detection, anti-Foxp3 antibody and Foxp3 Fix/Perm buffer were used according to the manufacturer’s instructions. The gating standards for PD-1 and Foxp3 were set using the isotype controls recommended by the manufacturer. A total of 30,000-50,000 CD3+ cells were collected with a BD FACS VERSE flow cytometer (BD Biosciences, San Jose, USA), and data were analyzed by Flowjo software (Flowjo LLC, USA) (38 (link)).
Pd 1 pe clone eh12.2h7
Manufactured by BioLegend
Sourced in United States
PD-1-PE (clone EH12.2H7) is a fluorochrome-conjugated antibody that binds to the programmed cell death protein 1 (PD-1). PD-1 is an inhibitory co-receptor expressed on activated T cells, B cells, and myeloid cells. This antibody can be used for the identification and characterization of PD-1-expressing cells in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Vδ2 percp clone b6, by BioLegend (1 mentions)
Facsverse flow cytometer, by BD (1 mentions)
Cd45ra bv510 clone hi100, by BioLegend (1 mentions)
Cxcr5 bv421 clone j252d4, by BioLegend (1 mentions)
Canto 2 instrument, by BD (1 mentions)
Cd8 pe cy7 clone sk1, by BioLegend (1 mentions)
Hla dr fitc clone l243, by BioLegend (1 mentions)
Cd3 apc cy7 clone hit3a, by BioLegend (1 mentions)
2 protocols using pd 1 pe clone eh12.2h7
1
Characterization of PD-1 and Foxp3 Expression in AML
2
Comprehensive Immune Profiling of PBMC Samples
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Peripheral blood mononuclear cells (PBMC) were isolated from fresh venous blood using Ficoll density gradient. PBMC samples were stained with the following antibodies: CD3-APC-Cy7 (clone HIT3a), CD3-BV510 (clone OKT3), CD4-BV421 (clone OKT4), CD8-PE-Cy7 (clone SK1), CD45RA-BV510 (clone HI100), CCR7-APC (clone G043H7), CD27-FITC (clone MT271), HLA-DR-FITC (clone L243), CXCR5-BV421 (clone J252D4), PD-1-PE (clone EH12.2H7), CXCR3-BV510 (clone G025H7), CCR4-PerCP-Cy5.5 (clone L291H4), CCR6-PE (clone G034E3), CD25-APC (clone BC96), CD127-FITC (clone A019D5), Perforin-PE-Cy7 (dG9), Granzyme B-AF647 (GB11) were purchased from Biolegend (San Diego, CA); CD4-percp (clone SK3), CD38-APC (clone HIT2), Tim-3-PE (clone 7D3), GNLY-AF488 (clone RB1) were obtained from BD Biosciences (San Diego, CA). Granzyme B, Perforin and GNLY were measured de novo, without prior stimulation with PMA/ionomycin. BD Canto II instrument was used for FACS and the data was analyzed using FlowJo software V10 (Tree star Inc. Ashland, OR).
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