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Anti cd45ra apc cy7 hi100

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Anti-CD45RA-APC-Cy7 (HI100) is a mouse anti-human monoclonal antibody that binds to the CD45RA antigen. It is conjugated with APC-Cy7 fluorescent dye.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Anti ccr7 bv785, by BioLegend (1 mentions) Anti cd3 bv510 hit3a, by BD (1 mentions) Nlvpmvatv, by JPT Peptide Technologies (1 mentions) Anti cd8 percp cy5.5 sk1, by BD (1 mentions) Lsr 2, by BD (1 mentions) Anti cd40 hb14, by Miltenyi Biotec (1 mentions) Anti pe magnetic beads, by Miltenyi Biotec (1 mentions)

4 protocols using anti cd45ra apc cy7 hi100

1

Phenotyping CMV-specific T Cells

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Cryopreserved PBMCs were stained with cell trace violet (CTV; Thermo Fisher) (20 minutes, RT), and then stimulated with 1 μg/mL epitope peptide (NLVPMVATV; JPT Peptide Technologies) for 6 days at 37°C in 500 μL of 10% fetal bovine serum/RPMI 1640 medium (WELGENE). Cells were labeled with HLA-A∗0201-restricted CMVpp65 pentamer (HLA-A2∗0201: CMVpp56495, ProImmune). Next, surface molecule staining, dead cell staining, fixation, permeabilization, and intracellular molecule staining were performed using the following antibodies: anti-CD3-BV510 (HIT3a), anti-CD8-Percp-Cy5.5 (SK1; BD Biosciences), anti-CD19-PE-eFluor610 (HIB19), anti-CD14-PE-eFluor610 (61D3), anti-Ki-67-PE-Cy7 (20Raj1; Thermo Fisher), anti-CD45RA-APC-Cy7 (HI100), and anti-CCR7-BV785 (G043H7; BioLegend), and a Foxp3 Staining Buffer Set. Data were collected using an LSR II instrument.
Kim S., Lee S.W., Koh J.Y., Choi D., Heo M., Chung J.Y., Lee B.H., Yang S.H., Sung Y.C., Lee H., Shin E.C, & Park S.H. (2022). A single administration of hIL-7-hyFc induces long-lasting T-cell expansion with maintained effector functions. Blood Advances, 6(23), 6093-6107.
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2

CMVpp65-Specific T Cell Activation Assay

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Cryopreserved PBMCs were labeled with HLA-A∗0201-restricted CMVpp65 pentamer (10 minutes, RT) and then stimulated with 1 μg/mL epitope peptide (NLVPMVATV) (4 hours, 37°C) in 200 μL/well of 10% fetal bovine serum/RPMI 1640 medium in 96-well round-bottom plates. Next, the cells were stained with interferon gamma and tumor necrosis factor (TNF) catch reagents (Miltenyi Biotec) (55 minutes, 37°C) on a rotator, and then with FITC-labeled anti-IFN-γ and PE-labeled anti-TNF detection reagents (Miltenyi Biotec) (10 minutes, 4°C). Next, cells were relabeled with pentamer (10 minutes, RT), and stained with anti-CD3-BV510 (HIT3a), anti-CD8-BV711 (SK1; BD Biosciences), anti-CD45RA-APC-Cy7 (HI100), and anti-CCR7-BV785 (G043H7; BioLegend). Data were collected using an LSR II instrument.
Kim S., Lee S.W., Koh J.Y., Choi D., Heo M., Chung J.Y., Lee B.H., Yang S.H., Sung Y.C., Lee H., Shin E.C, & Park S.H. (2022). A single administration of hIL-7-hyFc induces long-lasting T-cell expansion with maintained effector functions. Blood Advances, 6(23), 6093-6107.
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3

Comprehensive T Cell Phenotyping

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T cells collected from duplicate wells were combined and stained with appropriate antibody conjugates for 30 minutes at 4°C, then washed and resuspended in 30 nM DAPI prepared in PBS. Cell populations were subsequently analyzed by flow cytometry (BD LSR II).
Antibody conjugates, Alexa Flour 647–anti-CD8 (RPA-T8), PE/Cy7–anti-CD4 (OKT4), Alexa Flour 700–anti-CD25 (BC96), FITC–anti-CD38 (HIT2), PE–anti–PD-1 (EH12.2h7), PerCP-Cy5.5–anti–LAG-3 (11C3C65), Dazzle–anti-CD27 (0323), and APC/Cy7–anti-CD45RA (HI100), and appropriate isotype controls were purchased from BioLegend. DAPI was bought from Thermo Fisher Scientific.
Matus E.I., Sparkes A, & Gariépy J. (2020). A soluble activator that favors the ex vivo expansion of CD8+CD27+ T cells. JCI Insight, 5(22), e141293.
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4

Antigen-specific CD4+ T cell isolation

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Fresh PBMCs (10–20 × 106 cells, final concentration 10 × 106 cells/ml) were stimulated for 3 h at 37°C with 0.6 nmol/ml PepTivator C. albicans MP65 (pool of 15-amino-acid-length peptides with 11 amino acid overlap, Miltenyi Biotec) in RPMI + 5% human serum in the presence of 1 μg/ml anti-CD40 (HB14, Miltenyi Biotec). After stimulation, the PBMCs were labeled with PE-conjugated anti-CD154 (5C8, Miltenyi Biotec) and enriched with anti-PE magnetic beads (Miltenyi Biotec) (41 (link)). After enrichment, the cells were stained with PerCP-Cy5.5 anti-CD4 (RPA-T4, Biolegend), AlexaFluor700 anti-CD3 (SK7, Biolegend), and APC-Cy7 anti-CD45RA (HI100, Biolegend), and the antigen-specific T cells were gated as CD3+CD4+CD45RA−CD154+ single live cells for single cell sorting.
Attaf N., Cervera-Marzal I., Dong C., Gil L., Renand A., Spinelli L, & Milpied P. (2020). FB5P-seq: FACS-Based 5-Prime End Single-Cell RNA-seq for Integrative Analysis of Transcriptome and Antigen Receptor Repertoire in B and T Cells. Frontiers in Immunology, 11, 216.
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