Multicolor flow cytometric analysis of human Langerin (CD207) expression by monocyte-derived Langerhans cells. Peripheral blood monocytes were cultured with Recombinant Human GM-CSF (Cat. No. 550068; 100 ng/ml), IL-4 (Cat. No. 554605; 20 ng/ml), and TGF-β (Corning; Cat. No. 354039; 40 ng/ml) proteins for 3 days followed by an additional 3-day culture with GM-CSF and TGF-β. The cells were washed and stained with APC Mouse Anti-Human CD1a antibody (Cat. No. 559775/561755) and BD Horizon™ Fixable Viability Stain 450 (Cat. No. 562247). After washing, the cells were fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were subsequently washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Panel) or PE Mouse Anti-Human Langerin (CD207) antibody (Cat. No. 564727; Right Panel). Flow cytometric contour plots showing the correlated expression of Langerin (CD207) [or Ig Isotype control staining] versus CD1a were derived from gated events with the forward and side light-scatter, and viability stain characteristics of intact cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System.
.png){kind=spacer}
Multicolor flow cytometric analysis of human Langerin (CD207) expression by monocyte-derived Langerhans cells. Peripheral blood monocytes were cultured with Recombinant Human GM-CSF (Cat. No. 550068; 100 ng/ml), IL-4 (Cat. No. 554605; 20 ng/ml), and TGF-β (Corning; Cat. No. 354039; 40 ng/ml) proteins for 3 days followed by an additional 3-day culture with GM-CSF and TGF-β. The cells were washed and stained with APC Mouse Anti-Human CD1a antibody (Cat. No. 559775/561755) and BD Horizon™ Fixable Viability Stain 450 (Cat. No. 562247). After washing, the cells were fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were subsequently washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Panel) or PE Mouse Anti-Human Langerin (CD207) antibody (Cat. No. 564727; Right Panel). Flow cytometric contour plots showing the correlated expression of Langerin (CD207) [or Ig Isotype control staining] versus CD1a were derived from gated events with the forward and side light-scatter, and viability stain characteristics of intact cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System.
Product Details
BD Pharmingen™
CLC4K; CLEC4K; C-type lectin domain family 4 member K
Human (QC Testing)
Mouse IgG1, λ
Human Langerin Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested), Flow cytometry (Tested During Development)
5 µl/test
AB_2738920
Aqueous buffered solution containing BSA and ≤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
- 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.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- An isotype control should be used at the same concentration as the antibody of interest.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Fixation and Permeabilization Solution RUO
Size
125 mL
Cat No.
554722
Perm/Wash Buffer RUO
Size
100 mL
Cat No.
554723
Fixable Viability Stain 450 RUO
Size
0.1 mg
Cat No.
562247
PE Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
554680
564727 Rev. 1
Antibody Details
2G3
The 2G3 monoclonal antibody specifically binds to Langerin which is also known as CD207 or C-type lectin domain family 4 member K (CLEC4K). CD207 is a 48 kDa type II transmembrane glycoprotein that belongs to the C-type lectin family. CD207 is selectively expressed by Langerhans cells, an immature dendritic cell type that is found in the epidermis and mucosal epithelia, and by some other dendritic cell subsets. CD207 is expressed on the cell surface membrane and by intracellular Birbeck granules whose formation depends on CD207. CD207 binds to certain surface carbohydrates that are expressed by microbes. It appears to play a role in the uptake of these microbes for subsequent processing and microbial antigen presentation to T cells.
564727 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
564727 Rev.1
Citations & References
View product citations for antibody "564727" on CiteAb
Development References (5)
-
Flamar AL, Zurawski S, Scholz F, et al. Noncovalent assembly of anti-dendritic cell antibodies and antigens for evoking immune responses in vitro and in vivo. J Immunol. 2012; 189(5):2645-2655. (Clone-specific: Functional assay). View Reference
-
Igyarto BZ, Haley K, Ortner D, et al. Skin-resident murine dendritic cell subsets promote distinct and opposing antigen-specific T helper cell responses. Immunity. 2011; 35(2):260-272. (Immunogen: ELISA, Flow cytometry, Fluorescence microscopy, Functional assay, Immunofluorescence, In vivo exacerbation). View Reference
-
Mayumi N, Watanabe E, Norose Y, et al. E-cadherin interactions are required for Langerhans cell differentiation. Eur J Immunol. 2013; 43(1):270-280. (Biology). View Reference
-
Valladeau J, Ravel O, Dezutter-Dambuyant C, et al. Langerin, a novel C-type lectin specific to Langerhans cells, is an endocytic receptor that induces the formation of Birbeck granules. Immunity. 2000; 12(1):71-81. (Biology). View Reference
-
de Witte L, Nabatov A, Pion M, et al. Langerin is a natural barrier to HIV-1 transmission by Langerhans cells. Nat Med. 2007; 13(3):367-371. (Biology). View Reference
564727 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.