Anti cd45ro

Manufactured by BD

Sourced in United States

Anti-CD45RO is a monoclonal antibody used for the identification and enumeration of T cells expressing the CD45RO isoform of the CD45 antigen. CD45RO is a marker of memory T cells. This antibody can be used in flow cytometry applications.

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Lab products found in correlation

Anti cd4, by BD (11 mentions) Anti cd45ra, by BD (10 mentions) Anti cd3, by BD (7 mentions) Anti cd8, by BD (7 mentions) Anti hla dr, by BD (4 mentions) Anti cd4 rpe, by BD (4 mentions) Anti cd8 rpe, by BD (4 mentions) Anti cd4 fitc anti cd25 pe, by BD (4 mentions) Anti cd3 fitc anti cd56 rpe, by Agilent Technologies (4 mentions) Anti pd 1, by BD (4 mentions) Anti cd45, by BD (4 mentions) Anti cd56, by BD (3 mentions) Anti cd3 fitc, by Agilent Technologies (3 mentions) Fc500, by Beckman Coulter (3 mentions) Cxp analysis software, by Beckman Coulter (3 mentions) Anti ccr7, by BD (3 mentions) Anti cd28, by BD (3 mentions) Facsdiva software, by BD (2 mentions) Isotype matched irrelevant antibodies, by BD (2 mentions) Lsrfortessa, by BD (2 mentions)

Spelling variants of this product

Anti-CD45RO-FITC (13 citations) Anti-CD45RO-APC (10 citations) Anti–CD45RO-PE-Cy7 (7 citations) PE-conjugated anti-CD45RO (3 citations) APC-labeled anti-CD45RO (3 citations) Anti-CD45RO-PE (UCHL1, (2 citations) APC Mouse Anti-Human CD45RO (2 citations) Anti-CD45RO-BUV395 (clone UCHL1, (2 citations) Anti-CD45RO-BV421 (2 citations) FITC-conjugated anti-CD45RO (UCHL1; (2 citations)

21 protocols using anti cd45ro

Comprehensive Immune Cell Profiling

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Freshly prepared PBMCs were used. Subpopulations of T cells, B cells, natural killer (NK) cells and antigen-presenting cells (APC) were characterized by surface staining with fluorescence labelled anti-CD3, anti-CD4, anti-CD5, anti-CD8, anti-CD14, anti-CD16, anti-CD19, anti-CD25, anti-CD27, anti-CD38, anti-CD45RA, anti-CD45RO, anti-CD56 and anti-HLA-DR (BD Bioscience); anti-BDCA1, anti-BDCA2, anti-BDCA3, anti-BDCA4 and anti-slan (Miltenyi Biotec). Negative controls included directly labeled or unlabeled isotype-matched irrelevant antibodies (BD Biosciences). Freshly prepared CSF cells were used directly for FACS analysis. Fluorescence-labeled antibodies for surface staining were used as follows: anti-CD3, anti-CD4, anti-CD8, anti-CD14, anti-CD16, anti-CD19, anti-CD25, anti-CD27, anti-CD45RA, anti-CD45RO, anti-CD56 and anti-HLA-DR (BD Bioscience); anti-BDCA1 and anti-slan (Miltenyi Biotec). All cells were measured on a LSR-Fortessa (BD Biosciences) and evaluated by FACS-Diva Software (BD Bioscience).

Jin M., Günther R., Akgün K., Hermann A, & Ziemssen T. (2020). Peripheral proinflammatory Th1/Th17 immune cell shift is linked to disease severity in amyotrophic lateral sclerosis. Scientific Reports, 10, 5941.

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Phenotypic Analysis of T-cell Subsets

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T-lymphocyte purification and their subsequent activation were analyzed by flow cytometry (BD FACSCanto™; Becton Dickinson Immunocytometry Systems, San José, CA, USA). Cells were stained using the following monoclonal antibodies conjugated with fluorescein isothiocyanate (FITC) or phycoerythrin (PE): anti-CD4 (CD4⁺ T-lymphocytes), anti-CD25α (activated CD4⁺ T-lymphocytes), anti-CD45RA (naïve CD4⁺ T-lymphocytes), and anti-CD45RO (memory CD4⁺ T-lymphocytes) following the manufacturer’s recommendations (BD Biosciences Pharmingen, San José, CA, USA). Isotype-matched control monoclonal antibodies were used to determine the negative cell populations.

ALVAREZ C., BENÍTEZ A., ROJAS L., PUJOL M., CARVAJAL P., DÍAZ-ZÚÑIGA J, & VERNAL R. (2015). Differential expression of CC chemokines (CCLs) and receptors (CCRs) by human T lymphocytes in response to different Aggregatibacter actinomycetemcomitans serotypes. Journal of Applied Oral Science, 23(6), 580-590.

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Immunophenotyping and Cytokine Analysis

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Freeze-dried BCG (Chengdu Institute of Biological Products, Chengdu, China) was reconsitituted in a 0.9% sodium chloride solution to a concentration of 1 mg/mL prior to use. Sonicatd M.tb protein antigen (TB-Ag) was generously provided by Prof. Baiqing Li (Department of Immunology, Bangbu Medical College, Anhui Key Laboratory of Infection and Immunity, Bangbu, China). The following mAbs were used for phenotypic and intracellular cytokine analysis: phycoerythrin (PE)-labeled anti-CD3, anti-CD14, anti-CD56, anti-CD45RA, anti-IL-17, fluorescein isothiocyanate (FITC)-labeled anti-CD3, anti-CD45RA, anti-CD45RO, anti-granzyme B, Phycoerythrin-Cy7 (PE-cy7)-labeled anti-CD69, anti-CD56, allophycocyanin (APC)-labeled anti-CD3, anti-IFN-γ anti-IL-22 were obtained from BD Pharmingen (San Jose, CA, USA). FITC-labeled anti-CD16 was purchased from Biolegend (CA, USA). PE-labeled anti-NKG2D were obtained from R&D Systems (MN, USA), respectively.

Fu X., Yu S., Yang B., Lao S., Li B, & Wu C. (2016). Memory-Like Antigen-Specific Human NK Cells from TB Pleural Fluids Produced IL-22 in Response to IL-15 or Mycobacterium tuberculosis Antigens. PLoS ONE, 11(3), e0151721.

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Comprehensive Immune Profiling of Checkpoint Blockade

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Tissue staining was performed retrospectively on a subset of 86 patients with IHC-accessible tissue. All of these patients were treated with an immunological checkpoint blockade. Of all patients treated with immunological checkpoint block, 61 were excluded due to inaccessible pathology or insufficient IHC samples (eg, samples obtained by fine needle aspiration biopsy). Tissue staining was performed on PD-L1 (tumor), PD-L1 (Immunomicroenvironment), CD3, CD4, CD8, CD25, and CD56. The primary antibody against PD-L1 (SP263, Ventana) was used according to the manufacturer instructions. primary antibody of anti-CD3-FITC/anti-CD56-RPE (Dako), anti-CD3-FITC (fluorescein isothiocyanate), anti-CD4-RPE, anti-CD8-RPE, anti-CD45RO, and anti-CD4-FITC/anti-CD25-PE (BD Biosciences) were used. The sample was read and explained by the attending pathologist. The data report the percentage of cells stained for each particular marker.

Ren F., Zhao T., Liu B, & Pan L. (2019). Neutrophil–lymphocyte ratio (NLR) predicted prognosis for advanced non-small-cell lung cancer (NSCLC) patients who received immune checkpoint blockade (ICB). OncoTargets and therapy, 12, 4235-4244.

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Quantifying Immune Cell Phenotypes

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Peripheral venous blood was obtained from each patient before surgery. Most of the patients were collected at the second day of admission. Whole blood (100 ml) was incubated in the dark with primary antibody at 4°C for 15 min. anti-CD3-FITC/anti-CD56-RPE (Dako), anti-CD3-FITC (fluorescein isothiocyanate), anti-CD4-RPE, anti-CD8-RPE, anti-CD45RO and anti-CD4-FITC/anti-CD25-PE (BD Biosciences) were used. After hemolysis for 10 min, samples were centrifuged for 10 min at 1500 rpm at room temperature, and then washed twice in PBS and subjected to flow cytometric analysis. Three-color flow cytometric analysis was performed to determine cell phenotypes using an FC500 (Beckman–Coulter) and CXP analysis software (Beckman–Coulter). Lymphocytes were gated by forward scatter versus side scatter. Analysis was set to collect 5000 gated events.

Xiang Z.J., Hu T., Wang Y., Wang H., Xu L, & Cui N. (2020). Neutrophil–lymphocyte ratio (NLR) was associated with prognosis and immunomodulatory in patients with pancreatic ductal adenocarcinoma (PDAC). Bioscience Reports, 40(6), BSR20201190.

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Phenotypic Analysis of T-cell Subsets

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Multiparametric Flow Cytometry Protocol

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The following monoclonal antibodies were used for flow cytometry analysis and cell sorting; anti-CD8 (TONBO Biosciences), anti-CD4 (BD Biosciences), anti-CD45RA (BioLegend), anti-CD45RO (BD Biosciences). LIVE/DEAD fixable yellow dead cell stain kit (Thermo Fisher Scientific). Recombinant human IL-2 was obtained through the AIDS Research and Reference Reagent Program (Division of AIDS, National Institute of Allergy and Infectious Diseases). Recombinant human IL-7 and IL-15 were purchased from BioLegend.

Ogura H., Preston-Hurlburt P., Perdigoto A.L., Amodio M., Krishnaswamy S., Clark P., Yu H., Egli D., Fouts A., Steck A.K, & Herold K.C. (2018). Identification and analysis of islet antigen specific CD8+ T cells with T cell libraries. Journal of immunology (Baltimore, Md. : 1950), 201(6), 1662-1670.

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Multimodal Analysis of Cellular Adhesion Proteins

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Anti-FAK, anti-ALCAM-PE, anti-ICAM1-PE, anti-VCAM1-PE, anti-VE cadherin, anti-von Willebrand (vWF) factor, anti-CHS1 were purchased from Abcam (Cambridge, MA, USA). OX124, mouse anti-human monoclonal antibody against HS, was a kind gift from Dr. Marion Brown, Oxford, UK. Biotin-labeled Anti-CD18 KIM127 was purchased from Exploratory Research Cell Tech Therapeutics Ltd. (Slough, UK). Anti-phospho-SLP-76; pTyr128, anti-VE-cadherin were purchased from Cell signaling Technologies (Danvers, MA, USA). Anti-HIF-1, anti-pZAP-70-FITC, anti-pZAP-70-PE, anti- LAG3, anti-TIM3, anti-PD-1, eFluor 670, anti-CD45-APC, anti-CHS-PerCP, anti-CCR7, anti-CD45RO were purchased from BD biosciences (Franklin Lakes, NJ, USA). Anti-Talin-1 and anti-Vinculin were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Goat anti-human Fc-PE was purchased from Millipore (Billerica, MA, USA). Alexa Fluor (AF) labelled secondary anti-mouse, anti-goat anti-rabbit antibodies, streptavidin AF 488 antibody; Texas Red Phalloidin and AF488 Phalloidin were purchased from Life Technologies (Carlsbad, CA, USA). Anti-Fab DyLight 488 was purchased from Jackson ImmunoResearch (Suffolk, UK)

Samaha H., Pignata A., Fousek K., Ren J., Lam F., Stossi F., Dubrulle J., Salsman V.S., Krishnan S., Hong S.H., Baker M.L., Shree A., Gad A.Z., Shum T., Fukumura D., Byrd T.T., Mukherjee M., Marrelli S.P., Orange J.S., Joseph S.K., Sorensen P.H., Taylor M.D., Hegde M., Mamonkin M., Jain R.K., El-Naggar S, & Ahmed N. (2018). A Homing System Targets Therapeutic T-cells to Brain Cancer. Nature, 561(7723), 331-337.

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Multicolor Flow Cytometry of Lymphocytes

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Peripheral venous blood was obtained from each patient at various time points. Whole blood (100 mL) was incubated with primary antibody in the dark for 15 mins at 4°C. anti-CD3-FITC/anti-CD56-RPE (Dako), anti-CD3-FITC (fluorescein isothiocyanate), anti-CD4-RPE, anti-CD8-RPE, anti-CD45RO and anti-CD4-FITC/anti-CD25-PE (BD Biosciences). After 10 mins of hemolysis, the samples were centrifuged at 1,500 rpm for 10 mins at room temperature, then washed twice in PBS and subjected to flow cytometry analysis. Three-color flow cytometry analysis was performed using FC500 (Beckman-Coulter) and CXP analysis software (Beckman-Coulter) to determine the cell phenotype. Lymphocytes are gated by forward scatter and side scatter. The analysis will collect 5,000 gated events.

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Multiparametric Flow Cytometry Analysis

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Cells were stained with saturating amounts of various fluorescent-labeled antibody combinations including anti-EpCAM (EBA-1 clone), anti-CD45 (HI30 clone), anti-CD3e (UCHT1 clone), anti-CD56 (B159 clone), anti-CD4 (SK3 clone), anti-CD8 (SK1 clone), anti-CD25 (M-A215 clone), anti-CD107a (H4A3 clone), anti-CD45RO (REA611 clone), anti-NKG2A (REA110 clone), anti-NKG2D (BAT221 clone), anti-CD137 (4B4–1 clone), anti-CD16 (3G8 clone), anti-HLA-E (3D12 clone), Annexin V and co-stained with DAPI (all from BD Biosciences or Miltenyi). Cells were analyzed with the Attune NxT flow cytometer (Life Technologies) and further analyses were performed with FlowJo software (Tree Star).

Courau T., Bonnereau J., Chicoteau J., Bottois H., Remark R., Assante Miranda L., Toubert A., Blery M., Aparicio T., Allez M, & Le Bourhis L. (2019). Cocultures of human colorectal tumor spheroids with immune cells reveal the therapeutic potential of MICA/B and NKG2A targeting for cancer treatment. Journal for Immunotherapy of Cancer, 7, 74.

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