Anti cd45ra ecd

Manufactured by Beckman Coulter

Sourced in United States, United Kingdom

Anti-CD45RA-ECD is a laboratory reagent used for the detection and identification of cells expressing the CD45RA antigen. It is a monoclonal antibody conjugated with the fluorescent dye ECD (Phycoerythrin-Texas Red).

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Lab products found in correlation

Anti pd 1 pe, by BD (1 mentions) Anti cd25 pe cy7, by BD (1 mentions) Facsaria flow cytometer, by BD (1 mentions) Anti cd3 brilliant violet 570, by BioLegend (1 mentions) Anti ccr7 brilliant violet 421, by BioLegend (1 mentions) Anti lag 3 apc, by R&D Systems (1 mentions) Anti ctla 4 pe cy5, by BD (1 mentions) Anti tim3 percp efluor710, by Thermo Fisher Scientific (1 mentions) Anti cd4 brilliant violet 510, by BioLegend (1 mentions) Anti cd8a apc cy7, by BD (1 mentions) Anti cd25 pe cy7, by Thermo Fisher Scientific (1 mentions) Anti cd80 fitc, by Thermo Fisher Scientific (1 mentions) Anti pd l1 pe, by Thermo Fisher Scientific (1 mentions) Anti cd8 pe cy5, by Thermo Fisher Scientific (1 mentions) Lsrii cytometer, by BD (1 mentions) Anti cd3 alexa 700, by BD (1 mentions) Anti cd20 ecd, by Beckman Coulter (1 mentions) Anti eomes efluor 660, by Thermo Fisher Scientific (1 mentions) Anti cd27 apc cy7, by BioLegend (1 mentions) Anti pd 1 fitc, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

CD45RA-ECD (10 citations) Anti-CD45RA (4 citations) Anti-CD45RA–ECD (clone 2H4LDH11LDB9; (2 citations) ECD-conjugated anti-CD45RA (clone 2H4) (2 citations)

4 protocols using anti cd45ra ecd

Comprehensive T-cell Phenotyping by Flow Cytometry

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One million T-cells were stained with the following Ab cocktail: anti-CD3 Brilliant Violet 570 (Biolegend, CA, USA), anti-CD4 Brilliant Violet 510 (Biolegend, CA, USA), anti-CD8a APC-Cy7 (BD Biosciences, CA, USA), anti-4-1BB FITC (eBioscience, CA, USA), anti-CD127 APC-AF700 (Beckman Coulter, CA, USA), anti-CD45RA ECD (Beckman Coulter, CA, USA), anti-CCR7 Brilliant Violet 421 (Biolegend, CA, USA), anti-LAG-3 APC (R&D Systems, Minneapolis, MN), anti-CD25 PE-Cy7 (BD Biosciences, CA, USA), anti-CTLA-4 PE-Cy5 (BD Biosciences, CA, USA), anti-TIM3 Percp-eFluor710 (eBioscience, CA, USA) and anti-PD-1 PE (BD Biosciences, CA, USA). After 15 min, T-cells were washed with PBS–0.1% FBS and analyzed using a FACSAria flow cytometer (BD Biosciences, Stockholm, Sweden); data analysis was performed using FlowJo software.

Liu Z., Poiret T., Persson O., Meng Q., Rane L., Bartek J J.r., Karbach J., Altmannsberger H.M., Illies C., Luo X., Harvey-Peredo I., Jäger E., Dodoo E, & Maeurer M. (2017). NY-ESO-1- and survivin-specific T-cell responses in the peripheral blood from patients with glioma. Cancer Immunology, Immunotherapy, 67(2), 237-246.

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Comprehensive PBMC Immunophenotyping Protocol

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Peripheral blood mononuclear cells (PBMCs) were stained in four panels containing anti-CD3-PacificBlue (PacBlue) (antibodies from BD Biosciences unless otherwise indicated), anti-CD3-Alexa700, anti-CD25-PE-Cy7, anti-CD38-PE, anti-HLA-DR-PE-Cy7, anti-CCR5-APC, anti-CD123-PerCP-Cy5.5, anti-CD16-PacBlue, anti-CD80-FITC, anti-CD83-PE, anti-CD86-APC, anti-PD1-FITC, anti-PD-L1-PE, anti-HLA class I-APC, anti-CD69-APC-Cy7; anti-CD4-Qdot655, anti-CD8-PE-Cy5.5, anti-CD14-Qdot605 (Invitrogen); anti-CD45RA-ECD, anti-CD127-PE, anti-HLA-DR-ECD, anti-CD20-ECD (Beckman Coulter); anti-CD11c-Alexa700 (eBioscience); and anti-CD27-APC-Cy7 (BioLegend). A staining reagent for dead cells (Invitrogen Aqua Live/Dead Fixable Stain) was included. Cells were then washed and fixed in PBS containing 1% paraformaldehyde or permeabilized using a FOXP3 Fix/Perm kit (BioLegend) according to the manufacturer's instructions, intracellularly stained with anti-Ki67-Alexa 488 (BD Biosciences), anti-FOXP3-PacBlue, anti-T-Bet-BV711 (BioLegend), and anti-Eomes-eFluor 660 (eBioscience), fixed, and analyzed with a LSR II cytometer (Becton Dickinson) and FlowJo.

Méndez-Lagares G., Lu D., Chen C., Terrault N., Segal M.R., Khalili M., Monto A., Shen H., Manos M.M., Lanier L.L., Ryan J.C., McCune J.M, & Hartigan-O'Connor D. (2017). Memory T-cell proliferation before HCV therapy predicts antiviral immune responses and treatment success. Journal of immunology (Baltimore, Md. : 1950), 200(3), 1124-1132.

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Multiparameter Immune Cell Analysis

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We used the following fluorescence-conjugated monoclonal antibodies and staining reagents: anti-CD3-v500, anti-CD8-allophyocyanine (APC)-H7, anti-CD27-phycoerythrine (PE), interleukin 2 (IL-2)-fluorescein isothiocyanate (FITC), tumor necrosis factor α (TNF-α)-Alexa 700, and CD107a-APC (all BD Biosciences, Oxford, United Kingdom); anti-CD4-peridinin chlorophyll (PerCP), anti-interferon γ (IFN-γ) PE-cyanine 7, anti-CD154 Pacific-Blue (BioLegend, Cambridge, United Kingdom), anti-CD45RA-ECD (Beckman Coulter, United Kingdom), and Yellow live-dead stain (Invitrogen, Paisley, United Kingdom).

Bajwa M., Vita S., Vescovini R., Larsen M., Sansoni P., Terrazzini N., Caserta S., Thomas D., Davies K.A., Smith H, & Kern F. (2016). Functional Diversity of Cytomegalovirus–Specific T Cells Is Maintained in Older People and Significantly Associated With Protein Specificity and Response Size. The Journal of Infectious Diseases, 214(9), 1430-1437.

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Isolation and Sorting of CD4+ T Cell Subsets

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All participants underwent leukaphereses performed as outpatients. PBMC were isolated and viably cryopreserved. Frozen PBMC were thawed and CD4 T cells enriched with the EasySep™ Human CD4 + T Cell Negative Selection Enrichment Kit (Stemcell). Cells were stained with Live/Dead Fixable Aqua (Life Technologies) and the following monoclonal antibodies cocktail: anti-CD3-FITC, anti-CD4-AlexaFluor700, anti-CCR7-PE-Cyanine7, anti-CD27-APC, anti-HLA-DR-APC H7, anti-CD57-Brilliant Violet 421, and anti-CD95-PE (Becton Dickinson) as well as anti-CD45RA-ECD (Beckman Coulter). HLA-DR- CD4 + T cell subpopulations were sorted on a FACS ARIA II flow cytometer (BD Biosciences) at >97% purity. Dry pellets were snap-frozen at −80 °C. Flow cytometry data were analyzed on FACSDiva v8.0.1 (BD Biosciences) and FlowJo v8.7 (Tree Star). Sorting schema is provided in Supplementary Fig. 2.

Reeves D.B., Bacchus-Souffan C., Fitch M., Abdel-Mohsen M., Hoh R., Ahn H., Stone M., Hecht F., Martin J., Deeks S.G., Hellerstein M.K., McCune J.M., Schiffer J.T, & Hunt P.W. (2023). Estimating the contribution of CD4 T cell subset proliferation and differentiation to HIV persistence. Nature Communications, 14, 6145.

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