Anti cd45ro

Manufactured by BioLegend

Sourced in United States

Anti-CD45RO is a laboratory reagent used to identify and study T cells that express the CD45RO isoform of the CD45 surface protein. It is commonly used in flow cytometry, immunohistochemistry, and other immunological research applications.

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Lab products found in correlation

Anti ccr4, by BioLegend (2 mentions) Anti hla dr, by BioLegend (1 mentions) Anti cd27, by BioLegend (1 mentions) Anti cd38, by BioLegend (1 mentions) Anti nkg2d, by BioLegend (1 mentions) Anti pd 1, by BioLegend (1 mentions) Anti tim 3, by BioLegend (1 mentions) Anti cd226, by BioLegend (1 mentions) Live dead fixable near ir dye, by Thermo Fisher Scientific (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Cd4 specific microbeads, by Miltenyi Biotec (1 mentions) Anti ccr6, by Thermo Fisher Scientific (1 mentions) Anti cxcr3, by BD (1 mentions) Anti il1r1, by R&D Systems (1 mentions) Anti cd45ra, by Thermo Fisher Scientific (1 mentions) Anti ccr10, by R&D Systems (1 mentions) Cd14 specific microbeads, by Miltenyi Biotec (1 mentions) Anti cd4, by Thermo Fisher Scientific (1 mentions) Il 23, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

PE anti-CD45RO (5 citations) PE-conjugated anti-CD45RO (4 citations) Anti-CD45RO PECy7 (4 citations) Anti-CD45RO (UCHL1, (3 citations) Anti-CD45RO FITC (2 citations) Anti-CD45RO Pacific Blue (2 citations) Anti-B220/CD45RO (2 citations) PE anti-human CD45RO (2 citations) Anti-CD45RO APC (2 citations) Anti-CD45RO-PerCP-Cy5.5 (2 citations)

4 protocols using anti cd45ro

Vδ2+ T Cell Differentiation Upon DC Coculture

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Peripheral blood lymphocytes (PBLs) from HSCT recipients were cocultured with pamidronate-pretreated autologous or allogeneic (refers to third-party healthy subjects) DCs at the ratio of 2:1, at a final of 3 × 10⁵ cells/well in 96-well plates. In some experiments, the neutralization antibody anti-NKG2D (10 μg/ml, BioLegend, USA) was used for blocking NKG2D during coculture. After 7 and 14 days of coculture, the percentages and differentiation and activation phenotypes of Vδ2⁺ T cells were detected by flow cytometry. Different combinations of monoclonal antibodies allowed identifying the differentiation profile of Vδ2⁺ T cells by the expression of CD45RO and CD27 (Naive: CD45RO⁻CD27⁺, Central Memory (CM): CD45RO⁺CD27⁺, Effector Memory (EM): CD45RO⁺CD27⁻ and terminal differentiation (TD): CD45RO⁻CD27⁻). For cell differentiation and activation assay, the cultured cells were stained with fluorochrome-labeled anti-CD27, anti-CD45RO, anti-HLA-DR, anti-CD38, and anti-NKG2D antibodies (BioLegend, USA).

Wang X., Liu J., Gao H., Mo X.D., Han T., Xu L.P., Zhang X.H, & Huang X.J. (2018). Dendritic Cells Are Critical for the Activation and Expansion of Vδ2+ T Cells After Allogeneic Hematopoietic Transplantation. Frontiers in Immunology, 9, 2528.

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Multiparametric Flow Cytometry of PBMCs

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For multiparametric flow cytometry analysis, PBMC were stained and enriched with the matched MHC class I or class II tetramer. To exclude dead cells in the subsequent analysis, PBMC were stained with the LIVE/DEAD™ Fixable Near-IR dye (Thermo Fisher, Germany) according to the manufacturer’s protocol. PBMC were stained with appropriate fluorochrome-conjugated surface antibodies, including anti-CD3 (OKT3), anti-CD4 (RPA-T4) anti-CD45RO (UCHL1), anti-CCR7 (G043H7), anti-CD226 (DX11), anti-TIGIT (A15153G), anti-BTLA (MIH26), anti-LIGHT (115520), anti-Ceacam1 (ASL-32), anti-Tim-3 (F38-2E2), anti-PD-1 (EH12.2H7), anti-OX40 (ACT-35), anti-CD14 (63D3), anti-CD19 (HIB19) (from Biolegend, Koblenz or BD Biosciences, Heidelberg, Germany) for 20 min at RT in the dark. After surface staining, cells were washed once with PBS and were then resuspended in 0.5% paraformaldehyde.

Ackermann C., Smits M., Woost R., Eberhard J.M., Peine S., Kummer S., Marget M., Kuntzen T., Kwok W.W., Lohse A.W., Jacobs T., Boettler T, & Schulze zur Wiesch J. (2019). HCV-specific CD4+ T cells of patients with acute and chronic HCV infection display high expression of TIGIT and other co-inhibitory molecules. Scientific Reports, 9, 10624.

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Isolation and Culture of Human Memory T Cells

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Samples were obtained from healthy donors participating in the Benaroya Research Institute Immune Mediated Disease Registry. Informed consent was obtained from all subjects according to IRB approved protocols at Benaroya Research Institute. CD4⁺CD25⁻ cells were enriched from PBMCs by positive selection with CD4-specific microbeads (Miltenyi Biotec). Memory cell subsets were sorted to over 97% purity as CD4+CD45RA-CD45RO+CD127+CD25− using anti-CD45RA (eBioscience), anti-CD45RO (Biolegend), anti-CD127 (BD Horizon), anti-CD25 (Biolegend) and anti-CD4 (Invitrogen). Antibodies used for sorting of memory cell subsets were: anti-CCR6 (eBioscience); anti-CCR10 (R&D Systems), anti-CCR4 (Biolegend), anti-CXCR3 (BD Pharmingen), anti-IL1R1 (R&D Systems). CD14+ monocytes were isolated from PBMCs by positive selection with CD14-specific microbeads (Miltenyi Biotec). Cells were cultured in RPMI 1640 medium supplemented with 2 mM glutamine, 1% (vol/vol) nonessential amino acids, 1% (vol/vol) sodium pyruvate, penicillin (50 U/ml), streptomycin (50 µg/ml) (all from Invitrogen) and 5% heat-inactivated human serum. In some experiments, recombinant human IL-1β (10 ng/ml; R&D Systems), IL-12 (10 ng/ml; R&D Systems) or IL-23 (25 ng/ml; eBioscience) were added to the culture.

Duhen T, & Campbell D.J. (2014). IL-1β promotes the differentiation of polyfunctional human CCR6+CXCR3+ Th1/17 cells that are specific for pathogenic and commensal microbes. Journal of immunology (Baltimore, Md. : 1950), 193(1), 120-129.

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Characterization of Murine and Human Immune Cells

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Both OTX015 and JQ1 (Stratech Scientific, UK) were dissolved in DMSO to make a stock solution of 10 mM. Aliquots were kept at -80 o C for up to 6 months and diluted with appropriate culture media for in vitro use. Murine antibodies for cell culture (anti-CD3, anti-CD28, anti-IL-4, anti-IFN-) and flow cytometry (anti-IL-17, anti-IFN-) were purchased from eBioscience (USA) and human antibodies (anti-IL-17, anti-IFN-) were purchased from Biolegend (USA) or eBioscience (USA). Sorting antibodies (anti-CD3, anti-CD4, anti-CD44, anti-CD62L for murine and anti-CD45RA, anti-CD45RO, anti-CCR7, anti-CCR6, anti-CCR4, anti-CXCR3 for human) were purchased from Biolegend (USA).
Recombinant murine IL-6 and IL-12 were purchased from PeproTech (USA) and recombinant human TGF- was purchased from R&D systems (USA).

Hu X., Schewitz-Bowers L.P., Lait P.J.P., Copland D.A., Stimpson M.L., Li J.J., Liu Y., Dick A.D., Lee R.W.J, & Wei L. (2018). The Bromodomain and Extra-Terminal Protein Inhibitor OTX015 Suppresses T Helper Cell Proliferation and Differentiation. Current molecular medicine, 18(9).

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