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Multicolor flow cytometric analysis of CCL22 expression in GM-CSF-stimulated adherent PBMC. Adherent peripheral blood mononuclear cells (PBMC) were cultured (3 days at 37°C) with Recombinant Human GM-CSF (Cat. No. 550068). BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724) was added for the last 3.5 hours of culture. The cells were harvested and stained with FITC Mouse Anti-Human CD206 antibody (Cat. No. 551135) and BD Horizon™ Fixable Viability Stain 660 (FVS660; Cat. No. 564405) in serum-free buffer. After fixation with BD Cytofix™ Fixation Buffer (Cat. No. 554655), the cells were permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723), and stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Human CCL22 antibody (Cat. No. 565950; Right Plot). Two-color flow cytometric contour plots showing the correlated expression of CD206 versus CCL22 (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact, viable (FVS660-negative) cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

Multiparameter flow cytometric analysis of CCL22 expression in Raji cells. Cells from the human Raji (Burkitt's B cell lymphoma, ATCC CCL-86) cell line were cultured with BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) for 3.5 hours. The cells were harvested and stained with BD Horizon™ Fixable Viability Stain 660 (FVS660) in serum-free buffer. The cells were washed, fixed with BD Cytofix™ Fixation Buffer, and then permeabilized with BD Perm/Wash™ Buffer, and stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Human CCL22 antibody (Cat. No. 565950; Right Plot). Two-parameter flow cytometric contour plots showing the correlated expression of CCL22 (or Ig Isotype control staining) versus side light-scatter (SSC) signals were derived from gated events with the forward and side light-scatter characteristics of intact, viable (FVS660-negative) Raji cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
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BD Pharmingen™ PE Mouse Anti-Human CCL22

BD Pharmingen™ PE Mouse Anti-Human CCL22
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Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
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The T51-719 monoclonal antibody specifically recognizes C-C motif chemokine 22 (CCL22), which is also known as Macrophage-derived chemokine (MDC), Stimulated T-cell chemotactic protein 1 (STCP-1), or Small-inducible cytokine A22 (SCYA22). CCL22 is produced by macrophages, dendritic cells, B cells, and NK cells. CCL22 binds to and signals through the cell surface receptor, CCR4. It attracts chronically activated T cells, including cutaneous lymphocyte antigen (CLA)-positive T cells, Th2-like cells, and regulatory T cells into inflammatory sites. CCL22 also attracts NK cells, monocytes/macrophages, and dendritic cells.

Development References (8)
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Chang Ms, McNinch J, Elias C, et al. Molecular cloning and functional characterization of a novel CC chemokine, stimulated T cell chemotactic protein (STCP-1) that specifically acts on activated T lymphocytes.. J Biol Chem. 1997; 272(40):25229-37. (Biology). View Reference
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Ferenczi K, Fuhlbrigge RC, Pinkus J, Pinkus GS, Kupper TS. Increased CCR4 expression in cutaneous T cell lymphoma.. J Invest Dermatol. 2002; 119(6):1405-10. (Biology). View Reference
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Godiska R, Chantry D, Raport CJ, et al. Human macrophage-derived chemokine (MDC), a novel chemoattractant for monocytes, monocyte-derived dendritic cells, and natural killer cells.. J Exp Med. 1997; 185(9):1595-604. (Biology). View Reference
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Imai T, Chantry D, Raport CJ, et al. Macrophage-derived chemokine is a functional ligand for the CC chemokine receptor 4. J Biol Chem. 1998; 273(3):1764-1768. (Biology). View Reference
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Lin L, Nonoyama S, Oshiba A, Kabasawa Y, Mizutani S. TARC and MDC are produced by CD40 activated human B cells and are elevated in the sera of infantile atopic dermatitis patients.. J Med Dent Sci. 2003; 50(1):27-33. (Biology). View Reference
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Nakayama T, Hieshima K, Nagakubo D, et al. Selective induction of Th2-attracting chemokines CCL17 and CCL22 in human B cells by latent membrane protein 1 of Epstein-Barr virus.. J Virol. 2004; 78(4):1665-74. (Biology). View Reference
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Robertson MJ. Role of chemokines in the biology of natural killer cells.. J Leukoc Biol. 2002; 71(2):173-83. (Biology). View Reference
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Vulcano M, Albanesi C, Stoppacciaro A, et al. Dendritic cells as a major source of macrophage-derived chemokine/CCL22 in vitro and in vivo.. Eur J Immunol. 2001; 31(3):812-22. (Biology). View Reference
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