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RB545 Rat Anti-Mouse CD223 (LAG-3)
RB545 Rat Anti-Mouse CD223 (LAG-3)
Multiparameter flow cytometric analysis using BD OptiBuild™ RB545 Rat Anti-Mouse CD223 antibody (Cat. No. 756645; Right Plot) on viable C57BL/6 mouse splenocytes stimulated by plate-bound Rat Anti-Mosue CD3e antibody for 3 days, with Autofluorescence Control (Left Plot). Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed using FlowJo™ v10.8 software.
RB545 Rat Anti-Mouse CD223 (LAG-3)
Multiparameter flow cytometric analysis using BD OptiBuild™ RB545 Rat Anti-Mouse CD223 antibody (Cat. No. 756645; Right Plot) on viable C57BL/6 mouse splenocytes stimulated by plate-bound Rat Anti-Mosue CD3e antibody for 3 days, with Autofluorescence Control (Left Plot). Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed using FlowJo™ v10.8 software.
Multiparameter flow cytometric analysis using BD OptiBuild™ RB545 Rat Anti-Mouse CD223 antibody (Cat. No. 756645; Right Plot) on viable C57BL/6 mouse splenocytes stimulated by plate-bound Rat Anti-Mosue CD3e antibody for 3 days, with Autofluorescence Control (Left Plot). Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed using FlowJo™ v10.8 software.
Multiparameter flow cytometric analysis using BD OptiBuild™ RB545 Rat Anti-Mouse CD223 antibody (Cat. No. 756645; Right Plot) on viable C57BL/6 mouse splenocytes stimulated by plate-bound Rat Anti-Mosue CD3e antibody for 3 days, with Autofluorescence Control (Left Plot). Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed using FlowJo™ v10.8 software.
Product Details
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BD OptiBuild™
Lag3; LAG-3; Lymphocyte-activation gene 3; Ly66
Mouse (Tested in Development)
Rat LEW, also known as Lewis IgG1, κ
Mouse LAG3 fusion protein
Flow cytometry (Qualified)
0.2 mg/ml
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 to the dye under optimum conditions that minimize unconjugated dye and antibody.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  3. 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.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. An isotype control should be used at the same concentration as the antibody of interest.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  8. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
  9. Researchers should determine the optimal concentration of this reagent for their individual applications.
756645 Rev. 2
Antibody Details
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C9B7W

The C9B7W antibody monoclonal antibody specifically binds to an epitope in the D2 domain of CD223 (LAG3), the 70-kDa protein encoded by Lymphocyte-activation gene 3 (Lag3). A fusion protein consisting of the entire extracellular region of mouse LAG3 with mouse IgG1 was used as immunogen. CD223 is a type-I membrane protein with four extracellular Ig-like domains; it is structurally homologous to CD4; and, like CD4, it binds MHC class II molecules. However, unlike CD4, it is not expressed on resting human and mouse T lymphocytes. In the mouse, as previously described in the human, CD223 expression is upregulated on T lymphocytes (both CD4+ and CD8+) activated through the T-cell receptor (TCR) and on IL-2-activated NK (LAK) cells, and it is not detected on B cells, dendritic cells, or Phorbol 12-myristate 13-acetate (PMA)-stimulated splenocytes. Studies on human peripheral T lymphocytes suggest that CD223 associates with the TCR to downregulate TCR signaling. In contrast, in vivo and in vitro evaluations of vaccination protocols in mice suggest that CD223 promotes immune responses by activating antigen-presenting cells. Furthermore, NK cells of Lag3-/- mice display defects in their capacity to kill certain tumor cells. Mouse CD223 also has been demonstrated to contribute to the suppressor function of T regulatory cells and the C9B7W antibody has been shown to inhibit this function in vitro and in vivo. Therefore, CD223 appears to play complex roles in the regulation of immune responses. Although the C9B7W antibody is unable to block the binding of MHC class II-IgG2a fusion protein to CD223, it is able to block the CD223-mediated inhibition of IL-2 production by a T-cell hybridoma responding to antigen.

756645 Rev. 2
Format Details
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RB545
The BD Horizon RealBlue™ 545 (RB545) fluorochrome is part of the BD family of blue (488-nm) laser dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 496-nm and an emission maximum (Em Max) at 545-nm. Driven by BD innovation, RB545 can be used on spectral cytometers and is designed to be excited by the blue (488-nm) laser with minimal excitation by the 561-nm yellow-green laser. RB545 has minimal spillover into yellow-green detectors and a brightness level similar to FITC.  Given its unique emission max, on a spectral instrument RB545 can be used simultaneously with FITC and PE to provide an additional color off of the blue laser. Please ensure that your instrument configuration (lasers and optical filters) is appropriate for this dye.
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RB545
Blue 488 nm
496 nm
545 nm
756645 Rev.2
Citations & References
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756645 Rev. 2

 

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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.