Cd45ra bv786

Manufactured by BD

Sourced in United Kingdom

CD45RA-BV786 is a fluorescently-labeled monoclonal antibody that binds to the CD45RA isoform of the CD45 protein, which is expressed on the surface of certain types of immune cells. The BV786 fluorophore is used to label the antibody, allowing detection and analysis of CD45RA-positive cells by flow cytometry.

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Lab products found in correlation

Cd3 af700, by BD (1 mentions) Cd27 bv421, by BioLegend (1 mentions) Tcrαβ fitc, by BioLegend (1 mentions) P24 kc57 pe, by Beckman Coulter (1 mentions) Live dead aqua cell stain 405 nm, by Thermo Fisher Scientific (1 mentions) Facsaria cell sorter, by BD (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Sodium pyruvate, by Thermo Fisher Scientific (1 mentions) Cd25 pe, by BD (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Rapamycin, by Miltenyi Biotec (1 mentions) Cd4 apc fire750, by BioLegend (1 mentions) Human ril 2, by Miltenyi Biotec (1 mentions) Cd8 v500 rpa t8, by BD (1 mentions) X vivo 15 cell medium, by Lonza (1 mentions) Cd25 pe clone m a251, by BD (1 mentions)

2 protocols using cd45ra bv786

Comprehensive Immune Cell Profiling

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p24 KC57-PE was purchased from Beckman Coulter (Cat#6604667, Dilution 1/1000) and p24 28B7-APC was purchased from MediMabs (Cat#MM-0289-APC, Dilution 1/1000). CD3-AF700 (Clone: UCHT-1, Cat#557943, Dilution 1/100), CD45RA-BV786 (Clone: HI100, Cat#563870, Dilution 1/25) and CCR7-BB700 (Clone: 3D12, Cat#566437, Dilution 1/25) were purchased from BD Bioscience. CD8-FITC (Clone: BW135/80, Cat#130-113-719, Dilution 1/100) was purchased from Miltenyi/MACS. CD27-BV421 (Clone: O323, Cat#302823, Dilution 1/50), TCRαβ-FITC (Clone: IP26, Cat#306705, Dilution 1/50) and TCRγδ-PE-Cy7 (Clone: B1 Cat#331221, Dilution 1/50) were purchased from BioLegend. Live/Dead Aqua Cell Stain (405 nm) was purchased from ThermoFisher Scientific (Cat#L34957).

Gantner P., Pagliuzza A., Pardons M., Ramgopal M., Routy J.P., Fromentin R, & Chomont N. (2020). Single-cell TCR sequencing reveals phenotypically diverse clonally expanded cells harboring inducible HIV proviruses during ART. Nature Communications, 11, 4089.

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Isolation and Expansion of Human Regulatory T Cells

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Human ex vivo CD127^loCD25⁺CD4⁺ Tregs were flow sorted using a BD FACSAria cell sorter (BD Biosciences) after staining with: CD39-BB515 (clone Tu66, BD), CD25-PE (clone MA251, BD) or CD25-PE (BD), CD4 APCFire/750 (Clone A161A1, Biolegend) or CD4-ECD (Clone SFCI12T4D11, Beckman Coulter), CD73-BV421 (clone AD2, Biolegend), CD8 V500 (RPA-T8, BD), CD3-BV605 (SK7, Biolegend), CD127-AF647 (clone HIL-7R-M21, BD) or CD127-PEcy7 (eBioRDR5, ebioscience/Thermofisher) and CD45RA-BV786 (clone Hi100, BD), then expanded with 500–1000 U/mL human rIL2 (Miltenyi Biotec/Chiron) ±100 nM rapamycin (Miltenyi Biotec) and human Treg expansion kit anti-CD3/CD28 beads (Miltenyi Biotec/Invitrogen), using x-vivo 15 cell medium (Lonza) or RMPI-1640 medium supplemented with L-glutamine, penicillin-streptomycin (both PAA Laboratories, UK), sodium pyruvate (Gibco, UK) and 2–10% human AB pooled serum and media exchanged with fresh IL-2 every 3–5 days. For additional experiments, either memory Tregs (mTregs) were sorted as CD4⁺CD25⁺CD127^loCD45RA⁻, naïve Tregs were sorted as CD4⁺CD25⁺CD127^loCD45RA⁺ or CD39⁻CD73⁻ double negative Tregs were sorted as CD4⁺CD25⁺CD127^loCD39⁻CD73⁻. Beads were removed using MACSibead magnet (Milteny Biotec) prior to phenotyping or use in functional assays. Example sorts shown in Supplementary Figs. 1a and 2b).

Jarvis L.B., Rainbow D.B., Coppard V., Howlett S.K., Georgieva Z., Davies J.L., Mullay H.K., Hester J., Ashmore T., Van Den Bosch A., Grist J.T., Coles A.J., Mousa H.S., Pluchino S., Mahbubani K.T., Griffin J.L., Saeb-Parsy K., Issa F., Peruzzotti-Jametti L., Wicker L.S, & Jones J.L. (2021). Therapeutically expanded human regulatory T-cells are super-suppressive due to HIF1A induced expression of CD73. Communications Biology, 4, 1186.

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