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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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.
製品詳細
BD Pharmingen™
C-C motif chemokine 22; STCP-1; SCYA22; MDC; MDC(1-69); ABCD-1, DC/B-CK
Human (QC Testing)
Mouse IgG1, κ
Human CCL22 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_2739414
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation and Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
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.
関連製品
Stain Buffer (FBS) RUO
サイズ
500 mL
カタログ番号
554656
Stain Buffer (BSA) RUO
サイズ
500 mL
カタログ番号
554657
Fixation Buffer RUO
サイズ
100 mL
カタログ番号
554655
Perm/Wash Buffer RUO
サイズ
100 mL
カタログ番号
554723
Fixable Viability Stain 660 RUO
サイズ
100 µg
カタログ番号
564405
Protein Transport Inhibitor (Containing Monensin) RUO
サイズ
0.7 mL
カタログ番号
554724
565950 Rev. 1
抗体の詳細
T51-719
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.
565950 Rev. 1
フォーマットの詳細
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
565950 Rev.1
引用＆参考文献
View product citations for antibody "565950" on CiteAb
Development References (8)
-
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
565950 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.