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BV711 Rat Anti-Mouse CD223
BV711_CD223_vs_CD3_v1.png

Two-color flow cytometric analysis of CD223 expression on activated mouse splenocytes. C57BL/6 mouse splenocytes were stimulated in culture for 3 days with immobilized Purified NA/LE Hamster Anti-Mouse CD3e antibody (Cat. No. 553057). The cells were harvested, pre-incubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142) and then stained with FITC Hamster Anti-Mouse CD3e antibody (Cat. No. 553062/553061/561827) and either BD Horizon™ BV711 Rat IgG1, κ Isotype Control (Cat. No. 563283, Left Panel) or BD Horizon™ BV711 Rat Anti-Mouse CD223 antibody (Cat. No. 563179, Right Panel). Two-color flow cytometric dot plots showing the correlated expression patterns of CD223 (or Ig Isotype control staining) versus CD3e were derived from gated events with the forward and side light-scatter characteristics of viable activated splenocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

563179Image1.png

Two-color flow cytometric analysis of CD223 expression on activated mouse splenocytes. C57BL/6 mouse splenocytes were stimulated in culture for 3 days with immobilized Purified NA/LE Hamster Anti-Mouse CD3e antibody (Cat. No. 553057). The cells were harvested, pre-incubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142) and then stained with FITC Hamster Anti-Mouse CD3e antibody (Cat. No. 553062/553061/561827) and either BD Horizon™ BV711 Rat IgG1, κ Isotype Control (Cat. No. 563283, Left Panel) or BD Horizon™ BV711 Rat Anti-Mouse CD223 antibody (Cat. No. 563179, Right Panel). Two-color flow cytometric dot plots showing the correlated expression patterns of CD223 (or Ig Isotype control staining) versus CD3e were derived from gated events with the forward and side light-scatter characteristics of viable activated splenocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

BV711_CD223_vs_CD3_v1.png 563179Image1.png Red-Cap_Violet.png

Two-color flow cytometric analysis of CD223 expression on activated mouse splenocytes. C57BL/6 mouse splenocytes were stimulated in culture for 3 days with immobilized Purified NA/LE Hamster Anti-Mouse CD3e antibody (Cat. No. 553057). The cells were harvested, pre-incubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142) and then stained with FITC Hamster Anti-Mouse CD3e antibody (Cat. No. 553062/553061/561827) and either BD Horizon™ BV711 Rat IgG1, κ Isotype Control (Cat. No. 563283, Left Panel) or BD Horizon™ BV711 Rat Anti-Mouse CD223 antibody (Cat. No. 563179, Right Panel). Two-color flow cytometric dot plots showing the correlated expression patterns of CD223 (or Ig Isotype control staining) versus CD3e were derived from gated events with the forward and side light-scatter characteristics of viable activated splenocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Two-color flow cytometric analysis of CD223 expression on activated mouse splenocytes. C57BL/6 mouse splenocytes were stimulated in culture for 3 days with immobilized Purified NA/LE Hamster Anti-Mouse CD3e antibody (Cat. No. 553057). The cells were harvested, pre-incubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142) and then stained with FITC Hamster Anti-Mouse CD3e antibody (Cat. No. 553062/553061/561827) and either BD Horizon™ BV711 Rat IgG1, κ Isotype Control (Cat. No. 563283, Left Panel) or BD Horizon™ BV711 Rat Anti-Mouse CD223 antibody (Cat. No. 563179, Right Panel). Two-color flow cytometric dot plots showing the correlated expression patterns of CD223 (or Ig Isotype control staining) versus CD3e were derived from gated events with the forward and side light-scatter characteristics of viable activated splenocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Product Details
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BD Horizon™
Lag3; LAG-3; Lymphocyte-activation gene 3; Ly66
Mouse (QC Testing)
Rat LEW, also known as Lewis IgG1, κ
Mouse LAG3 fusion protein
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2737398
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 BD Horizon™ BV711 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV711 were removed.
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Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

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Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  7. BD Horizon Brilliant Violet 711 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
  8. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  9. Cy is a trademark of GE Healthcare.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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563179 Rev. 2
Antibody Details
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C9B7W
563179 Rev. 2
Format Details
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BV711
The BD Horizon Brilliant Violet™ 711 (BV711) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 407-nm and an acceptor dye with an emission maximum (Em Max) at 713-nm. BV711, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 710-nm (e.g., a 712/20-nm bandpass filter). The acceptor dye can be excited by the Red (628–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
BV711
407 nm
713 nm
563179 Rev.2
Citations & References
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View product citations for antibody "563179" on CiteAb

Development References (9)

  1. Baixeras E, Huard B, Miossec C, et al. Characterization of the lymphocyte activation gene 3-encoded protein. A new ligand for human leukocyte antigen class II antigens. J Exp Med. 1992; 176(2):327-337. (Biology). View Reference
  2. El Mir S, Triebel F. A soluble lymphocyte activation gene-3 molecule used as a vaccine adjuvant elicits greater humoral and cellular immune responses to both particulate and soluble antigens. J Immunol. 2000; 164(11):5583-5589. (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. Hannier S, Triebel F. The MHC class II ligand lymphocyte activation gene-3 is co-distributed with CD8 and CD3-TCR molecules after their engagement by mAb or peptide-MHC class I complexes. Int Immunol. 1999; 11(11):1745-1752. (Biology). View Reference
  5. Huang CT, Workman CJ, Flies D, et al. Role of LAG-3 in regulatory T cells. Immunity. 2004; 21(4):503-513. (Clone-specific: Flow cytometry, Functional assay, Inhibition, In vivo exacerbation). View Reference
  6. Huard B, Mastrangeli R, Prigent P, et al. Characterization of the major histocompatibility complex class II binding site on LAG-3 protein. Proc Natl Acad Sci U S A. 1997; 94(11):5744-5749. (Biology). View Reference
  7. Miyazaki T, Dierich A, Benoist C, Mathis D. Independent modes of natural killing distinguished in mice lacking Lag3. Science. 1996; 272(5260):405-408. (Biology). View Reference
  8. Prigent P, El Mir S, Dréano M, Triebel F. Lymphocyte activation gene-3 induces tumor regression and antitumor immune responses. Eur J Immunol. 1999; 29(12):3867-3876. (Biology). View Reference
  9. Workman CJ, Rice DS, Dugger KJ, Kurschner C, Vignali DA. Phenotypic analysis of the murine CD4-related glycoprotein, CD223 (LAG-3). Eur J Immunol. 2002; 32(8):2255-2263. (Immunogen: Flow cytometry, Functional assay, Inhibition). View Reference
View All (9) View Less
563179 Rev. 2

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.