Anti ccr4

Manufactured by BioLegend

Sourced in United States

Anti-CCR4 is a monoclonal antibody that binds to the CC chemokine receptor 4 (CCR4) expressed on the surface of certain immune cells. CCR4 is involved in the migration and activation of these cells. The primary function of this product is to detect and quantify CCR4-expressing cells in research applications.

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Lab products found in correlation

Close spelling variants of this product

Anti-CCR4-PE (3 citations) Anti-CCR4-PE-Cy7 (2 citations) PE-CY7-conjugated anti-CCR4 (2 citations) Anti-CCR4 BV421 (2 citations) Anti-CCR4-PE-Cy7 (clone L291H4, (2 citations)

5 protocols using anti ccr4

Comprehensive NK Cell Phenotyping

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The following fluorescently conjugated antibodies were used for phenotypic analysis of NK cells: anti-LFA-1 (363404); anti-CXCR3 (353720); anti-PD-1 (329906); anti-CD107a (328612); anti-NKP44 (325116); anti-CD158e1 (312706); anti-CD158d (347006); anti-CD27 (356412); anti-CCR4 (359412); anti-DNAM-1 (338316); anti-CD16 (302040); anti-Granzyme B (515406); anti-CD62L (304806); anti-CD69 (310910); anti-NKP80 (346706); anti-NKP30 (325210); anti-CD158f (341304); anti-CD158b (312612); anti-Tim-3 (345012); anti-CD94 (305504); anti-TIGIT (372706); anti-TRAIL (308206); anti-CD57 (322306); anti-CX3CR1 (341610); anti-NKG2D (320808); anti-Perforin (353310); anti-Ki67 (350504); anti-IFN-γ (506518); anti-CD94 (305506); anti-NKp46 (331916); anti-CTLA4 (369614); anti-CD96 (338416); anti-41BB (309818); anti-CD25 (356108) were purchased from Biolegend. anti-CD159a/NKG2A (FAB1059P) and anti-NKG2C (FAB138G) were purchased from R&D.
The following fluorescently conjugated antibodies were used to identify immune cell types in eNK or PBMC: anti-human Lineage Cocktail (348803); anti-CD56 (362550); anti-CD3 (300430); anti-CD33 (366620); anti-HLA-DR (307606); anti-CD14 (301836); anti-CD19 (302242); anti-CD11b (301322); anti-CD25 (356108); and anti-FOXP3 (320108) were purchased from Biolegend, San Diego, CA, USA.

Chen M., Li Y., Wu Y., Xie S., Ma J., Yue J., Lv R., Tian Z., Fang F, & Xiao W. (2021). Anti-Tumor Activity of Expanded PBMC-Derived NK Cells by Feeder-Free Protocol in Ovarian Cancer. Cancers, 13(22), 5866.

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Comprehensive Immunophenotyping of T Cells

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Cell culture media was RPMI 1640 (Mediatech Inc.) supplemented with 10% heat-inactivated FBS, 100 U/ml penicillin, 100 mg/ml streptomycin, 2 mM L-glutamine, and 50 mM 2-mercaptoethanol (Sigma-Aldrich). Fluorescent anti-CD4, anti-CD8α, anti-CD8β, anti-CD25, anti-CD44, anti-CD45, anti-CD45.1, anti-CD45.2, anti-CD62L, anti-CD103, anti-CCR4, anti-CCR6, anti-CCR7, anti-CCR-9, anti-TCRβ and anti-TCRγδ antibodies were purchased from Biolegend. Anti-CTLA-4, anti-PD-1, anti-GITR, anti-CD127, anti-IFNγ, anti-CXCR3 and anti-CXCR4 were purchased from BD Biosciences-Pharmingen. Anti-MHCII, anti-CD207, anti-Gr-1, anti-CD11b, and anti-Foxp3 staining kit were purchased from eBioscience. Murine recombinant IL-2, IL-6 and E-selection-FC were purchased from R&D Systems. BD Cytofix/Perm buffer was used for intracellular staining. Cells were run on a BD LSR II flow cytometer or a Beckman Coulter Gallios flow cytometer and analyzed by Flowjo (Flowjo LLC).

Zhang R., Borges C.M., Fan M.Y., Harris J.E, & Turka L.A. (2015). Requirement for CD28 in Effector Treg Differentiation, CCR6 induction, and Skin Homing. Journal of immunology (Baltimore, Md. : 1950), 195(9), 4154-4161.

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OSCC Secretome Effects on Teff and Tregs

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Sorted Teff and Tregs from healthy donors were activated with anti-CD3/CD28 beads (1:5 ratio) (Life Technologies) and 1000 UI IL-2 for 5 days a 37°C. Then, 100 uL of OSCC and control secretomes were added to 2x10⁵ Teff or 2x10⁵ Tregs (in 100uL) in XVIVO-15 serum-free medium 48h a 37°C. After the incubation, the supernatants were stored for further cytokine production measurement using the Cytokine Bead Array Th1/2/17 Kit (BD) and the cells were counted (CountBright Absolute Counting Beads), stained with Live/Dead dye (Life Technologies), anti-CXCR3, anti-CCR4, anti-CCR6, anti-CCR8, anti-PD-1 and anti-TIGIT (all BioLegend) and analyzed by flow cytometry. For the analysis of cells after secretome co-culture, cells were washed after co-culture with secretome and cultured in new media X-VIVO15 (LONZA) serum-free medium for 48 h at 37°C with anti-CD3/CD28 beads (1:5 ratio) (Life Technologies) and 1000 UI IL-2. After the incubation, the supernatants were stored for further cytokine production measurement and the cells were stained with Live/Dead dye (Life Technologies), anti-CCR6, anti-CCR8, anti-PD-1 and anti-TIGIT (all BioLegend).

Fraga M., Yáñez M., Sherman M., Llerena F., Hernandez M., Nourdin G., Álvarez F., Urrizola J., Rivera C., Lamperti L., Nova L., Castro S., Zambrano O., Cifuentes A., Campos L., Moya S., Pastor J., Nuñez M., Gatica J., Figueroa J., Zúñiga F., Salomón C., Cerda G., Puentes R., Labarca G., Vidal M., McGregor R, & Nova-Lamperti E. (2021). Immunomodulation of T Helper Cells by Tumor Microenvironment in Oral Cancer Is Associated With CCR8 Expression and Rapid Membrane Vitamin D Signaling Pathway. Frontiers in Immunology, 12, 643298.

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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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