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      RB705 Mouse Anti-Human LAG-3 (CD223)
      RB705 Mouse Anti-Human LAG-3 (CD223)

      Multicolor flow cytometric analysis of LAG-3 (CD223) expression on unstimulated (Top Plots) and stimulated (Bottom Plots) Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were cultured for 3 days with plate-bound Anti-Human CD3 (Cat. No. 567109/567108; 10 µg/mL for coating) and soluble Anti-Human CD28 (Cat. No. 555725; 1 µg/mL) antibodies, and Recombinant Human IL-2 Protein (Cat. No. 554603; 10 ng/mL). Unstimulated PBMC (from the same donor) and stimulated PBMC were stained with BD Horizon™ BUV395 Mouse Anti-Human CD8 (Cat. No. 563795; Left Plots), PE Mouse Anti-Human CD279 (PD-1) [Cat. No. 560795; Right Plots], and BD Horizon™ RB705 Mouse Anti-Human LAG-3 (CD223) [Cat. No. 570636/570722] antibodies. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. Bivariate pseudocolor density plots showing the correlated expression of LAG-3 (CD223) versus CD8 (Left Plots), or LAG-3 (CD223) versus CD279 (PD-1) [Right Plots] were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) unstimulated or stimulated lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.

      RB705 Mouse Anti-Human LAG-3 (CD223)

      Multicolor flow cytometric analysis of LAG-3 (CD223) expression on unstimulated (Top Plots) and stimulated (Bottom Plots) Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were cultured for 3 days with plate-bound Anti-Human CD3 (Cat. No. 567109/567108; 10 µg/mL for coating) and soluble Anti-Human CD28 (Cat. No. 555725; 1 µg/mL) antibodies, and Recombinant Human IL-2 Protein (Cat. No. 554603; 10 ng/mL). Unstimulated PBMC (from the same donor) and stimulated PBMC were stained with BD Horizon™ BUV395 Mouse Anti-Human CD8 (Cat. No. 563795; Left Plots), PE Mouse Anti-Human CD279 (PD-1) [Cat. No. 560795; Right Plots], and BD Horizon™ RB705 Mouse Anti-Human LAG-3 (CD223) [Cat. No. 570636/570722] antibodies. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. Bivariate pseudocolor density plots showing the correlated expression of LAG-3 (CD223) versus CD8 (Left Plots), or LAG-3 (CD223) versus CD279 (PD-1) [Right Plots] were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) unstimulated or stimulated lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.

      Product Details
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      BD Horizon™
      LAG3; CD223; FDC; Lymphocyte activation gene 3 protein; Protein FDC
      Human (QC Testing)
      Mouse IgG1, κ
      Human LAG-3 Recombinant Protein
      Flow cytometry (Routinely Tested)
      5 µl/test
      3902
      AB_3685914
      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.
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      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.

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      Product Notices

      1. 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.
      2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      3. 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).
      4. 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.
      5. 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.
      6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      7. An isotype control should be used at the same concentration as the antibody of interest.
      8. 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.
      9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      10. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
      11. For U.S. patents that may apply, see bd.com/patents.
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      570722 Rev. 1
      Antibody Details
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      T47-530

      The T47-530 specifically recognizes the Lymphocyte Activation Gene 3 (LAG-3) protein which is also known as, Protein FDC, or CD223. LAG-3 is a ~70 kDa type I transmembrane glycoprotein that belongs to the Ig superfamily and exhibits homology to CD4. LAG-3 is expressed on NK cells, regulatory T cells, and activated conventional T cells with higher expression found on CD8+ T cells compared with CD4+ T cells. LAG-3 is an activation induced cell surface molecule that like CD4, binds MHC class II molecules, but with much higher affinity. This may enable LAG-3 to act as a negative competitor of CD4 for MHC class II ligand binding. LAG-3 may associate with the TCR-CD3 complex to downregulate TCR signal transduction and T cell clonal expansion. In contrast, LAG-3-induced signaling may promote dendritic cell activation.

      570722 Rev. 1
      Format Details
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      RB705
      The BD Horizon RealBlue™ 705 (RB705) 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 707-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB705 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), RB705 can be used as an alternative to PerCP-Cy5.5 or BB700 and we recommend using an optical filter centered near 710-nm (e.g., a 695/40 or 710/50-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PerCP-Cy5.5. RB705 is on average brighter than PerCP-Cy5.5 and BB700, and has minimal spillover into Yellow-Green detectors.
      altImg
      RB705
      Blue 488 nm
      498 nm
      707 nm
      570722 Rev.1
      Citations & References
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      View product citations for antibody "570722" on CiteAb

      Development References (9)

      1. Al-Mterin MA, Murshed K, Elkord E. PD-1 expression, among other immune checkpoints, on tumor-infiltrating NK and NKT cells is associated with longer disease-free survival in treatment-naïve CRC patients.. Cancer Immunol Immunother. 2022. (Clone-specific: Flow cytometry). View Reference
      2. Casati C, Camisaschi C, Novellino L, et al. Human lymphocyte activation gene-3 molecules expressed by activated T cells deliver costimulation signal for dendritic cell activation. J Immunol. 2008; 180(6):3782-3788. (Biology). View Reference
      3. Hannier S, Tournier M, Bismuth G, Triebel F. CD3/TCR complex-associated lymphocyte activation gene-3 molecules inhibit CD3/TCR signaling. J Immunol. 1998; 161(8):4058-4065. (Biology). View Reference
      4. Huang CT, Workman CJ, Flies D, et al. Role of LAG-3 in regulatory T cells. Immunity. 2004; 21(4):503-513. (Biology). View Reference
      5. Moya R, Robertson HK, Payne D, Narsale A, Koziol J, Davies JD. A pilot study showing associations between frequency of CD4(+) memory cell subsets at diagnosis and duration of partial remission in type 1 diabetes.. Clin Immunol. 2016; 166-167:72-80. (Clone-specific: Flow cytometry). View Reference
      6. Murga-Zamalloa CA, Brown NA, Wilcox RA. Expression of the checkpoint receptors LAG-3, TIM-3 and VISTA in peripheral T cell lymphomas.. J Clin Pathol. 2020; 73(4):197-203. (Clone-specific: Flow cytometry). View Reference
      7. Toor SM, Murshed K, Al-Dhaheri M, Khawar M, Abu Nada M, Elkord E. Immune Checkpoints in Circulating and Tumor-Infiltrating CD4+ T Cell Subsets in Colorectal Cancer Patients.. Front Immunol. 2019; 10:2936. (Clone-specific: Flow cytometry). View Reference
      8. Triebel F, Hacene K, Pichon MF. A soluble lymphocyte activation gene-3 (sLAG-3) protein as a prognostic factor in human breast cancer expressing estrogen or progesterone receptors. Cancer Lett. 2006; 235(1):147-153. (Biology). View Reference
      9. Triebel F, Jitsukawa S, Baixeras E, et al. LAG-3, a novel lymphocyte activation gene closely related to CD4. J Exp Med. 1990; 171(5):1393-1405. (Biology). View Reference
      View All (9) View Less
      570722 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.