BD Horizon™ RB744 Mouse Anti-Human CD207 (Langerin)
Clone 2G3 (also known as AB5_8.2G3.6 ) (RUO)
Multicolor flow cytometric analysis of CD207 (Langerin) expression by monocyte-derived Langerhans cells. Human 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 BD Horizon™ BV421 Mouse Anti-Human CD1a antibody (Cat. No. 563938) and BD Horizon™ Fixable Viability Stain 510 (Cat. No. 564406). 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 BD Horizon™ RB744 Mouse IgG1, κ Isotype Control (Cat. No. 570519; Left Plot) or BD Horizon™ RB744 Mouse Anti-Human CD207 (Langerin) antibody (Cat. No. 570587/570675; Right Plot). The bivariate pseudocolor density plots showing the correlated expression of CD207(Langerin) [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 cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software.
Multicolor flow cytometric analysis of CD207 (Langerin) expression by monocyte-derived Langerhans cells. Human 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 BD Horizon™ BV421 Mouse Anti-Human CD1a antibody (Cat. No. 563938) and BD Horizon™ Fixable Viability Stain 510 (Cat. No. 564406). 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 BD Horizon™ RB744 Mouse IgG1, κ Isotype Control (Cat. No. 570519; Left Plot) or BD Horizon™ RB744 Mouse Anti-Human CD207 (Langerin) antibody (Cat. No. 570587/570675; Right Plot). The bivariate pseudocolor density plots showing the correlated expression of CD207(Langerin) [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 cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software.
Product Details
BD Horizon™
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
50489
Aqueous buffered solution containing ≤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 to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- 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.
- Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- Clones without a listed workshop have not been investigated in an HLDA workshop to receive a CD nomenclature. We use “CD” provisionally when our internal testing indicates that this clone binds to the same CD antigen as workshopped clones.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
RB744 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
570519
Fixation and Permeabilization Solution RUO
Size
125 mL
Cat No.
554722
Perm/Wash Buffer RUO
Size
100 mL
Cat No.
554723
Fixable Viability Stain 510 RUO
Size
100 µg
Cat No.
564406
570587 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.
570587 Rev. 1
Format Details
RB744
The BD Horizon RealBlue™ 744 (RB744) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 746-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB744 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), we recommend using an optical filter centered near 750-nm (e.g., a 750/60-nm bandpass filter).
RB744
Blue 488 nm
498 nm
746 nm
570587 Rev.1
Citations & References
View product citations for antibody "570587" on CiteAb
Development References (5)
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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
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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
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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
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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
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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
570587 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.