Anti cd4

Manufactured by Beckman Coulter

Sourced in United States

The Anti-CD4 is a laboratory reagent used for the identification and enumeration of CD4+ T-lymphocytes. It is designed for use with flow cytometry techniques.

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Lab products found in correlation

Anti cd8, by Beckman Coulter (3 mentions) Epics xl mcl, by Beckman Coulter (2 mentions) Anti hla dr, by Beckman Coulter (2 mentions) Anti cd45, by Beckman Coulter (2 mentions) Kaluza software, by Beckman Coulter (1 mentions) Gallios flow cytometer, by Beckman Coulter (1 mentions) Anti cd56, by Beckman Coulter (1 mentions) Anti cd3, by Beckman Coulter (1 mentions) Anti tcr vβ11, by Beckman Coulter (1 mentions) Anti cd80, by Beckman Coulter (1 mentions) Anti cd86, by Beckman Coulter (1 mentions) Anti cd3, by BD (1 mentions) Anti cd56, by BD (1 mentions) Anti cd19, by BD (1 mentions) Navios, by Beckman Coulter (1 mentions) Monensin, by Merck Group (1 mentions) 7 aad, by BD (1 mentions) Anti il 22, by Thermo Fisher Scientific (1 mentions) Anti il 9, by Thermo Fisher Scientific (1 mentions) Facscanto 2, by BD (1 mentions)

Spelling variants of this product

Anti-CD4-FITC (13 citations) Anti-CD4 ECD (10 citations) Anti-CD4-APC (9 citations) ), anti-CD4-PC5 (5 citations) Anti-CD4-APC-AlexaFluor750 (3 citations) Anti-human CD4 (APC, (2 citations) FITC-conjugated anti-CD4 (2 citations) Anti-CD4-RD1 (2 citations) Anti-CD4-PC5 anti-CD8-APC (2 citations) Anti-CD4-KO (2 citations)

6 protocols using anti cd4

Exosome Capture and Flow Cytometry Analysis

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Isolated exosomes were captured on aldehyde sulfate-coated latex beads (Invitrogen, Grand Island, NY, USA). Blocking of the beads with 0.5% w/v BSA was performed prior to capture. The non-specifically captured exosomes were detected with anti-CD9-FITC Ab (eBioSN4, eBioscience, San Diego, CA, USA) and mouse IgG1-FITC isotype (11-4714-42, eBioscience).
Cells to be used for flow cytometry were washed and incubated in the dark for 20 min with anti-CD4 (A07752, Beckman Coulter, Brea, CA, USA), anti-CD69 (555530, BD Biosciences, San Jose, CA, USA) or anti-CD154 (12-1548, eBioscience) Abs at room temperature. Isotypes were used for each experiment (mouse IgG1-κ-FITC, 555909, BD Biosciences and IgG1-κ-PE, 12-4714, eBioscience). Cells were again washed, re-suspended in flow buffer and analyzed using an EPICS XL-MCL or a Gallios flow cytometer (Beckman Coulter). At least 5x10⁴ events were collected, and the data were analyzed using the Kaluza software (Beckman Coulter).

Muller L., Hong C.S., Stolz D.B., Watkins S.C, & Whiteside T.L. (2014). Isolation of Biologically-Active Exosomes from Human Plasma. Journal of immunological methods, 411, 55-65.

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Multiparametric Flow Cytometry Analysis

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Surface phenotypes were determined using an Epics XL MCL (Beckman Coulter, Brea, CA, USA). The following monoclonal antibodies (mAbs) were purchased from Beckman Coulter: anti‐CD3, anti‐CD4, anti‐CD8, anti‐Vγ9TCR, anti‐CD14, anti‐CD25, anti‐CD45, anti‐CD54, anti‐CD56, anti‐HLA‐DR, anti‐CD40, anti‐CD80, anti‐CD86, anti‐CD11c, anti‐CD36, mouse immunoglobulin (Ig)G1, mouse IgG2 and mouse IgG2b mAbs. Anti‐HLA‐class 1 and anti‐CCR7 mAbs were purchased from Beckton Dickinson (San Jose, CA, USA) and R&D Systems (Minneapolis, MN, USA), respectively. Anti‐TCR Vα24TCR and anti‐TCR Vβ11 mAbs were purchased from Beckman Coulter (Villepinte, France). Anti‐human CD273 (PD‐L2) and CD274 (PD‐L1) mAbs were purchased form eBioscience (San Diego, CA, USA). Anti‐human CD152 (CTLA‐4) and CD279 (PD‐1) mAbs were purchased from BioLegend (San Diego, CA, USA). Anti‐FoxP3 mAb for intracellular staining was purchased from BD Biosciences (Tokyo, Japan). All mAbs were conjugated with fluorescein isothiocyanate (FITC), phycoerythrin (PE), antigen‐presenting cells (APC), extracellular domain (ECD), proprotein convertase (PC)5 or PC7. Z was purchased from Novartis Pharmaceuticals (Basel, Switzerland) and G from Funakoshi Co. Ltd (Tokyo, Japan).

Eiraku Y., Terunuma H., Yagi M., Deng X., Nicol A.J, & Nieda M. (2018). Dendritic cells cross‐talk with tumour antigen‐specific CD8+ T cells, Vγ9γδT cells and Vα24NKT cells in patients with glioblastoma multiforme and in healthy donors. Clinical and Experimental Immunology, 194(1), 54-66.

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Multiparameter Immunophenotyping of PBMCs

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Cell surface markers of peripheral blood mononuclear cells (PBMCs) were determined by immunofluorescent staining and flow cytometry (Navios, Beckman Coulter, Brea, CA, USA) using anti-CD3, anti-CD19, anti-CD16, and anti-CD56 from BD Biosciences; and anti-CD4 and anti-CD8 from Beckman Coulter.

Rechavi E., Lev A., Simon A.J., Stauber T., Daas S., Saraf-Levy T., Broides A., Nahum A., Marcus N., Hanna S., Stepensky P., Toker O., Dalal I., Etzioni A., Almashanu S, & Somech R. (2017). First Year of Israeli Newborn Screening for Severe Combined Immunodeficiency—Clinical Achievements and Insights. Frontiers in Immunology, 8, 1448.

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Intracellular Cytokine Staining of T Cells

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For intracellular cytokine staining cultured cells were restimulated with PMA (50 ng/mL, Sigma Aldrich) and Ionomycin (1 μg/mL, Invitrogen, Life technologies) in the presence of Monensin (2.5 mM, Sigma Aldrich) for 5 h. To measure the cell viability (>80%) the cells were stained with Annexin-V FITC and 7-AAD according to the manufacturer's protocol (BD Bioscience). The cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% saponin-containing buffer before washing and staining in different combinations with anti-CD4 (Beckman Coulter, Krefeld, Germany), anti-IL-17A, anti-IL-22, anti-IL-9, anti-IL-8, anti-AHR, anti-RORγt (ebioscience), and anti-IFN-γ (BD Bioscience). All dotplots and histograms shown refer to gated CD4⁺ T cells. Data were acquired on FACSCantoII (BD Bioscience) and were analyzed with FlowJo Software (Tree Star Inc., Ashland, OR, USA).

Gasch M., Goroll T., Bauer M., Hinz D., Schütze N., Polte T., Kesper D., Simon J.C., Hackermüller J., Lehmann I, & Herberth G. (2014). Generation of IL-8 and IL-9 Producing CD4+ T Cells Is Affected by Th17 Polarizing Conditions and AHR Ligands. Mediators of Inflammation, 2014, 182549.

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Evaluating Tumor Immune Response to Oncolytic Virus and BiTE

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Raji lymphoma cells were implanted intradermally into the right flanks of NSG mice (5 × 10⁶ cells). Mice bearing Raji subcutaneous tumor were treated intratumorally with PBS, OVV, OVV-CD19BiTE (2 × 10⁷ pfu/tumor), or blinatumomab (0.25 mg/kg). Twenty-four hours later, 1 × 10⁷ preactivated T cells were intravenously injected into treated mice. Tumors were harvested 3 days after the injection with forceps and surgical scissors and were weighed. They were then minced before incubation with Liberase (1.67 Wünsch U/mL) and DNase (0.2 mg/mL) in serum-free RPMI for 30 min at 37 °C. Cell suspensions were generated by mashing through a 70 μm nylon filter and then washed with complete RPMI. For tumor-infiltrating T-cells analysis, single-cell suspensions were generated and processed for surface labeling with anti-CD3, anti-CD45, anti-CD4, anti-CD8, anti-HLA-DR, anti-CD45RA, anti-CCR7, anti-CXCR3, and anti-CCR6 antibodies (Beckman). Live cells were distinguished from dead cells by using fixable dye eFluor506 (eBioscience). All flow data were acquired using either an LSRII flow cytometer (BD Biosciences). Data were analyzed with FlowJo software (Treestar).

Lei W., Ye Q., Hao Y., Chen J., Huang Y., Yang L., Wang S, & Qian W. (2022). CD19-targeted BiTE expression by an oncolytic vaccinia virus significantly augments therapeutic efficacy against B-cell lymphoma. Blood Cancer Journal, 12(2), 35.

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Isolation of Human Natural Killer Cells

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NK cells were isolated from human buffy coats of healthy donors obtained from the Etablissement Français du Sang. Informed consent was obtained from donors and experimental procedures were approved by the local institutional review committee. PBMCs were isolated by standard density gradient centrifugation on Ficoll-Hypaque (Eurobio). Mononuclear cells were separated from peripheral blood lymphocytes (PBLs) by centrifugation on a 50% Percoll solution (GE Healthcare). NK cells were purified from PBLs by immunomagnetic depletion using pan-mouse IgG Dynabeads (Thermo Fisher Scientific) with a cocktail of depleting monoclonal antibodies: anti-CD19 (4G7 hybridoma), anti-CD3 (OKT3 hybridoma, ATCC, Manassas, VA, USA), anti-CD4, anti-CD14 and anti-glycophorin A (all from Beckman Coulter). NK purity was >70% as assessed by CD56 labeling.

Perrin-Cocon L., Vidalain P.O., Jacquemin C., Aublin-Gex A., Olmstead K., Panthu B., Rautureau G.J., André P., Nyczka P., Hütt M.T., Amoedo N., Rossignol R., Filipp F.V., Lotteau V, & Diaz O. (2021). A hexokinase isoenzyme switch in human liver cancer cells promotes lipogenesis and enhances innate immunity. Communications Biology, 4, 217.

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