Anti cxcr4 pe

Manufactured by BD

Anti-CXCR4 PE is a fluorochrome-conjugated antibody that specifically binds to the CXCR4 chemokine receptor. It is designed for use in flow cytometry applications.

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CXCR4 (37 citations) CXCR4-PE (18 citations) Anti-CXCR4 (16 citations) PE-Cy5 mouse anti-human CXCR4 monoclonal antibody (3 citations) PE-labeled mouse anti-human CXCR4 (2 citations) Anti-CD184 (CXCR4)-PE (2 citations) PE-conjugated anti-CXCR4 (2 citations)

5 protocols using anti cxcr4 pe

Comprehensive Immune Cell Profiling

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Ab-stained cells were analyzed using FACScaliber (BD Biosciences, USA) CellQuest Software. Anti-CD4-PE, anti-CD4-APC, anti-CD8-FITC, anti-CD25-FITC, anti-CD44-PE, anti-CD62L-APC, anti-CD69-FITC, anti-CCR5-PE, anti-CXCR3-PE, and anti-CXCR4-PE, anti-IFNγ-BV421, anti-IL17A-PE, anti-Thy1.2-BV605 were purchased from BD Biosciences.

Park I., Son M., Ahn E., Kim Y.W., Kong Y.Y, & Yun Y. (2020). The Transmembrane Adaptor Protein LIME Is Essential for Chemokine-Mediated Migration of Effector T Cells to Inflammatiory Sites. Molecules and Cells, 43(11), 921-934.

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Absolute Enumeration of Tang Cells in SSc

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Trucount Tubes (BD Biosciences) were used for determining the absolute numbers of Tang cells in SSc patients and HC. Fresh whole peripheral blood (100 μl) was added into the Trucount Tube and then stained with anti-CD3 Pacific Blue, anti-CD8 PerCP-Cy5.5 (Miltenyi Biotec), anti-CD4 PE-Cy7, anti-CD31 FITC, anti-CXCR4 PE and anti-CD28 allophycocyanin (APC) antibodies (all from BD Biosciences). After 15 minutes, red blood cells were lysed with NH₄Cl and then cells were analyzed using a FACSCanto II flow cytometer (BD Biosciences) equipped with FACSDiva software (BD Biosciences). The absolute cell count was obtained multiplying the number of positive cell events by the number of Trucount Tube beads and subsequently dividing by the number of Trucount bead events.

Manetti M., Pratesi S., Romano E., Bellando-Randone S., Rosa I., Guiducci S., Fioretto B.S., Ibba-Manneschi L., Maggi E, & Matucci-Cerinic M. (2017). Angiogenic T cell expansion correlates with severity of peripheral vascular damage in systemic sclerosis. PLoS ONE, 12(8), e0183102.

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Immunophenotypic Analysis of Hematopoietic Cells

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Fresh leukapheresis products were stained with the following monoclonal antibodies (MoAbs): anti-CXCR4–PE, anti-CD49d–PE, anti-CD11a–PE, anti-CD34–FITC (BD Biosciences, Franklin Lakes, NJ) or corresponding isotype controls. Additionally, PB mononuclear cells (PBMNCs) were isolated by density gradient centrifugation, viably frozen, and stored in liquid nitrogen until use. Thawed PBMNCs were stained with anti-CD34–FITC (BD Biosciences), CD44–FITC, anti-CD38–PE, anti-CD34–PE (Beckman Coulter), and anti-CD133–APC MoAbs (Miltenyi Biotec, Bergisch-Gladbach, Germany). The percentage of cells expressing respective receptors and mean fluorescence intensity values in relation to isotype controls (MFIRs) were analyzed with a flow cytometer and its accompanying software (Gallios and Caluza, respectively, both Beckman Coulter).

Girbl T., Lunzer V., Greil R., Namberger K, & Hartmann T.N. (2014). The CXCR4 and adhesion molecule expression of CD34+ hematopoietic cells mobilized by “on-demand” addition of plerixafor to granulocyte–colony-stimulating factor. Transfusion, 54(9), 2325-2335.

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Immunophenotyping of T-cell Subsets

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Freshly isolated PBMCs were immunophenotyped for Treg number, FoxP3 expression (MFI) and CCR5/CxCR4 expression using fluorochrome-conjugated monoclonal antibodies (mAb): anti-CD4 PE/FITC, anti-CD25 PE-Cy7, anti-CCR5 PE, and anti-CxCR4 PE (BD Bioscience, San Jose, CA) for cell-surface markers in combination with intracellular Fork-head box protein 3 (FoxP3) mAb conjugated with Alexa488 (eBioscience, San Diego, CA). Samples were acquired into a flow cytometer (FACS Calibur, BD, USA) and analyzed using Cell-Quest (BD Bioscience)/FlowJo (Treestar) softwares. For further analysis, patients were categorized on the basis of level of surface CD25 expression on CD4⁺ T-cells viz: CD4⁺CD25^high (top 2% with high CD25 expression), CD4⁺CD25^intermediate (middle 15% with intermediate CD25 expression) and CD4⁺CD25^low/negative (lower 83% with very low or no expression of CD25) cells (Figure 1A & 1B). Absolute counts for different populations were calculated from the total lymphocyte count in the whole blood.

Toor J.S., Singh S., Sharma A, & Arora S.K. (2014). Mycobacterium tuberculosis Modulates the Gene Interactions to Activate the HIV Replication and Faster Disease Progression in a Co-Infected Host. PLoS ONE, 9(9), e106815.

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Flow Cytometry Analysis of Macrophage Differentiation

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Macrophages were differentiated in UpCell tissue culture plates (Thermo Scientific Nunc, Rochester, NY). After indicated times, cells were released from plates according to manufacturer’s protocol and stained in 0.5% BSA and 1 mM EDTA in PBS using the following antibodies, anti-CCR5 APC, anti-CXCR4 PE, anti-CD14 Pacific Blue, anti-CD16 PE-Cy7 (BD Biosciences, San Jose, CA), anti-CD4 FITC, anti-CD163 APC (ebioscience, San Diego CA), or anti-p24 PE antibody (KC57-RD1) (Beckman Coulter Indianapolis, IN). For intracellular p24 staining, cells were permeabilized and stained using the BD Cytofix/Cytoperm kit according to manufacturer’s instructions (BD Biosciences San Jose, CA). Flow cytometry was performed using a LSRII flow cytometer (BD Biosciences, San Jose, CA). Data were analyzed using Flow Jo software (Treestar, Ashland, OR).

Williams J.C., Appelberg S., Goldberger B.A., Klein T.W., Sleasman J.W, & Goodenow M.M. (2014). Δ9-tetrahydrocannabinol treatment during human monocyte differentiation reduces macrophage susceptibility to HIV-1 infection. Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology, 9(3), 369-379.

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