Skip to main content Skip to navigation
Hamburger Menu
Search icon
Order lookup icon Order lookup icon
Order Lookup
Location Selector Location Selector
United States (English)
Profile Icon Profile Icon
Sign-in/Register UserName
Products

Link Arrow
Solutions

Link Arrow
Discover & Learn

Link Arrow
Resources & Tools

Link Arrow
Support

Link Arrow
Close Menu

Dear Customer,  If you experience any text readability or display issues on our website, please update your web browser to the latest version

See More

Close
Website Feedback
Alert Icon
Alert icon
This product is the replacement for 744985.
569616Image1.png

Multicolor flow cytometric analysis of LAG-3 (CD223) expression on viable 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 Purified NA/LE Mouse Anti-Human CD3 (Cat. No. 567109; 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 (Top Plots) and stimulated PBMC (Bottom Plots) were stained with PE Mouse Anti-Human CD8 (Cat. No. 561950; Left Plots), BD Horizon™ BV421 Mouse Anti-Human CD279 (PD-1) [Cat. No. 562516/565935; Right Plots], and BD Horizon™ BV510 Mouse Anti-Human LAG-3 (CD223) [Cat. No. 569616] antibodies at 1.0 μg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) 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 (7-AAD-negative) unstimulated or stimulated lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

569616-4.png 569616Image1.png Green-Cap_Violet.png

Multicolor flow cytometric analysis of LAG-3 (CD223) expression on viable 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 Purified NA/LE Mouse Anti-Human CD3 (Cat. No. 567109; 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 (Top Plots) and stimulated PBMC (Bottom Plots) were stained with PE Mouse Anti-Human CD8 (Cat. No. 561950; Left Plots), BD Horizon™ BV421 Mouse Anti-Human CD279 (PD-1) [Cat. No. 562516/565935; Right Plots], and BD Horizon™ BV510 Mouse Anti-Human LAG-3 (CD223) [Cat. No. 569616] antibodies at 1.0 μg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) 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 (7-AAD-negative) unstimulated or stimulated lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Product Details
Down Arrow Up Arrow


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)
0.2 mg/ml
3902
AB_3685182
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 and unconjugated antibody and free dye were removed.
Show More blue down arrow Show Less blue up arrow

Recommended Assay Procedures

   BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to 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 CompBead to ensure that BD® CompBeads are appropriate for your specific cellular application.

   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).

Show More blue down arrow Show Less blue up arrow

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  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. BD Horizon Brilliant Violet 510 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
  7. 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.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. 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.
  10. For U.S. patents that may apply, see bd.com/patents.
Show More blue down arrow Show Less blue up arrow
569616 Rev. 1
Antibody Details
Down Arrow Up Arrow
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.

569616 Rev. 1
Format Details
Down Arrow Up Arrow
BV510
The BD Horizon Brilliant Violet™ 510 (BV510) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye with an excitation maximum (Ex Max) at 327-nm / 405-nm and an emission maximum (Em Max) at 512-nm. BV510, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 510-nm (e.g., a 525/50 bandpass filter). The dye can be excited by the UV (355-nm) laser resulting in cross-laser excitation and spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
BV510
327 nm, 405 nm
512 nm
569616 Rev.1
Citations & References
Down Arrow Up Arrow
View product citations for antibody "569616" 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
569616 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.