Cd45ro apc bv421

Manufactured by BD

CD45RO-APC/BV421 is a fluorescent-labeled antibody used to detect the presence of the CD45RO protein, which is a marker of memory T cells, in flow cytometry analysis.

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Lab products found in correlation

Cd62l pe, by BD (2 mentions) Cd62l pe cy7, by BioLegend (2 mentions) Cd4 pe fitc apc, by BD (2 mentions) 41bb apc pe, by BD (2 mentions) Cd8 pe cy7 fitc, by BD (2 mentions) Ox40 fitc pe cy7, by BD (2 mentions) Sh800s ma900 instrument, by Sony (2 mentions) Cd8a efluor 450, by Thermo Fisher Scientific (2 mentions) Cd3 alexa fluor 700, by BioLegend (2 mentions) Facscanto 3, by BD (2 mentions) Percp cy5, by Thermo Fisher Scientific (2 mentions) Cd3 apc cy7, by BD (2 mentions) Facsaria 2, by BD (2 mentions)

2 protocols using cd45ro apc bv421

T Cell Phenotyping and Sorting

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The antimouse TCRβ (mTCR)- PerCp-Cy5.5, CD8a-eFluor 450, were purchased from eBioscience. CD3-APC-Cy7, CD4- PE/FITC/APC, 41BB- APC/PE, CD8- PE-Cy7/FITC, OX40- FITC/PE-Cy7, CD62L-PE, and CD45RO-APC/BV421 were purchased from BD Biosciences. CD3-Alexa Fluor 700, CD62L-PE-Cy7, and the live/dead stains- DAPI/PI were purchased from BioLegend.
For FACS sample preparation, cells were harvested and washed with FACS buffer. Cells were resuspended in FACS buffer at a concentration of 1–50 × 10⁶ cells/mL and extracellular fluorescence-conjugated primary antibodies were added and mixed in. After a 20- to 60-minute incubation at 4°C, which was also protected from light exposure, the samples' cells were washed and resuspended with FACS buffer.
The flow cytometry assays were performed on FACSCanto I/II (BD Biosciences) and the acquired data were analyzed with FlowJo software (TreeStar). Cells were sorted to: Live/CD3⁺, separated for CD4⁺ or CD8⁺, T_EMRA/T_EM/T_CM based on sorting out CD62L⁺/CD45RO⁻ (naïve T cells). Cocultures were sorted for enrichment or into single reactive cells based on 41BB⁺/OX40⁺ (both double- and single-positive cells), Live/CD3⁺, separated for CD4⁺ or CD8⁺ by SH800S/MA900 instrument (Sony Biotechnology) or by FACSAria II (BD Biosciences).

Levin N., Paria B.C., Vale N.R., Yossef R., Lowery F.J., Parkhurst M.R., Yu Z., Florentin M., Cafri G., Gartner J.J., Shindorf M.L., Ngo L.T., Ray S., Kim S.P., Copeland A.R., Robbins P.F, & Rosenberg S.A. (2021). Identification and Validation of T-cell Receptors Targeting RAS Hotspot Mutations in Human Cancers for Use in Cell-based Immunotherapy. Clinical Cancer Research, 27(18), 5084-5095.

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Multiparametric Flow Cytometry Profiling of T Cell Subsets

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The anti-mouse TCRβ (mTCR)- PerCp-Cy5.5, CD8a-eFluor 450, were purchased from eBioscience. CD3-APC-Cy7, CD4- PE/FITC/APC, 4–1BB- APC/PE, CD8- PE-Cy7/FITC, OX40- FITC/PE-Cy7, CD62L-PE, and CD45RO-APC/BV421 were purchased from BD (Becton Dickinson Biosciences). CD3-Alexa Fluor 700, CD62L-PE-Cy7, and the live/dead stains- DAPI/PI were purchased from Biolegend.
For FACS sample preparation, cells were harvested and washed with FACS buffer. Cells were resuspended in FACS buffer at a concentration of 1e6–50e6 cells/ml and extracellular fluorescent-conjugated primary antibodies were added and mixed in. After a 20–60 min incubation at 4^oC, which was also protected from light exposure, the samples’ cells were washed and resuspended with FACS buffer.
The flow cytometry assays were performed on FACSCanto I/II (BD Biosciences) and the acquired data were analyzed with FlowJo software (TreeStar). Cells were sorted to: Live/CD3⁺, separated for CD4⁺ or CD8⁺, T_EMRA/ T_EM/T_CM based on sorting out CD62L⁺/CD45RO^- (T naïve cells). Cocultures were sorted for enrichment or into single reactive cells based on 41BB⁺/OX40⁺ (both double and single-positive cells), Live/CD3⁺, separated for CD4⁺ or CD8⁺ by SH800S/MA900 instrument (Sony Biotechnology) or by FACSAria II (BD).

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