Flow cytometric analysis of human CCL18 expression in cultured immature monocyte-derived dendritic cells or unstimulated peripheral blood monocytes. Immature monocyte-derived dendritic cells (MoDC; Left Panel) were generated from peripheral blood monocytes cultured in complete Iscove's Modified Dubelcco's Medium (IMDM) containing recombinant human IL-4 (Cat. No. 554605) and GM-CSF (Cat. No. 550068) for 5 days. Freshly isolated, peripheral blood monocyte (PB-M; Right Panel) cells served as control cells. Cells were fixed and permeabilized using BD Cytofix/Cytoperm™ Fixation/Permablization Kit reagents (Cat. No. 554714) followed by staining with PE Mouse anti-Human CCL18 (Cat. No. 561179). Histograms show the expression pattern of CCL18 (Solid line) or cellular autofluorescence (dotted line) on these two cell populations derived by gating events with the forward and side light-scatter characteristics of intact MoDC and PB-M. Flow cytometry was performed on a BD LSR™II Flow Cytometry System.
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Flow cytometric analysis of human CCL18 expression in cultured immature monocyte-derived dendritic cells or unstimulated peripheral blood monocytes. Immature monocyte-derived dendritic cells (MoDC; Left Panel) were generated from peripheral blood monocytes cultured in complete Iscove's Modified Dubelcco's Medium (IMDM) containing recombinant human IL-4 (Cat. No. 554605) and GM-CSF (Cat. No. 550068) for 5 days. Freshly isolated, peripheral blood monocyte (PB-M; Right Panel) cells served as control cells. Cells were fixed and permeabilized using BD Cytofix/Cytoperm™ Fixation/Permablization Kit reagents (Cat. No. 554714) followed by staining with PE Mouse anti-Human CCL18 (Cat. No. 561179). Histograms show the expression pattern of CCL18 (Solid line) or cellular autofluorescence (dotted line) on these two cell populations derived by gating events with the forward and side light-scatter characteristics of intact MoDC and PB-M. Flow cytometry was performed on a BD LSR™II Flow Cytometry System.
Product Details
BD Pharmingen™
C-C motif chemokine 18; Dendritic cell chemokine 1; MIP-4; PARC; AMAC-1
Human (QC Testing)
Mouse IgG1, κ
Human CCL18 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
6362
AB_10611716
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO
Preparation And Storage
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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Recommended Assay Procedures
Please see the Support Protocols for Intracellular Cytokine Staining on the BD Biosciences website. http://www.bdbiosciences.com/support/resources/cytokines/index.jsp
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- An isotype control should be used at the same concentration as the antibody of interest.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Fixation/Permeabilization Kit RUO
Size
250 Tests
Cat No.
554714
Recombinant Human IL-4 RUO
Size
5 µg
Cat No.
554605
Recombinant Human GM-CSF RUO
Size
10 µg
Cat No.
550068
561179 Rev. 1
Antibody Details
AZN-CK18
The AZN-CK18 monoclonal antibody specifically binds to the CC chemokine ligand 18 (CCL18), also known as Dendritic cell chemokine 1 (DC-CK1). CCL18 is expressed by dendritic cells and activated (eg, with LPS or cytokines including IL-4, IL-10, IL-13) monocytes and macrophages. CCL18 is a chemotactic factor that primarily attracts T and B lymphocytes (but not monocytes or granulocytes) towards dendritic cells and macrophages. This suggests that CCL18 likely plays roles in the development of both humoral and cell-mediated immune responses and may participate is certain inflammatory diseases, such as those affecting the joints and lungs.
561179 Rev. 1
Format Details
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
561179 Rev.1
Citations & References
Development References (3)
-
Lindhout E, Vissers JL, Hartgers FC, et al. The dendritic cell-specific CC-chemokine DC-CK1 is expressed by germinal center dendritic cells and attracts CD38-negative mantle zone B lymphocytes. J Immunol. 2001; 166(5):3284-3289. (Clone-specific: Immunohistochemistry). View Reference
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Schutyser E., Richmond A., and Van Damme J. Involvement of CC chemokine ligand 18(CCL18) in normal and pathological process. J Leukoc Biol. 2005; 78(1):14-26. (Biology). View Reference
-
van der Voort R, Kramer M, Lindhout E, et al. Novel monoclonal antibodies detect elevated levels of the chemokine CCL18/DC-CK1 in serum and body fluids in pathological conditions. J Leukoc Biol. 2005; 77(5):739-747. (Immunogen). View Reference
561179 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.
