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APC-R700 Mouse Anti-Human LAG-3 (CD223)
APC-R700 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. 555329; 10 µg/mL for coating) and soluble Anti-Human CD28 (Cat. No. 555725; 1 µg/mL) antibodies, and Human Recombinant IL-2 (Cat. No. 554603; 10 ng/mL). Unstimulated PBMC (from the same donor) and the stimulated PBMC were stained with PE Mouse Anti-Human CD8 (Cat. No. 555367/557086/561949/561950), BD Horizon™ BV421 Mouse Anti-Human CD279 (PD-1) (Cat. No. 562516/565935), and BD Horizon™ APC-R700 Mouse Anti-Human LAG-3 (CD223) (Cat. No. 565774/565775) antibodies. Two-color flow cytometric contour 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 unstimulated (Top Plots) or stimulated (Bottom Plots) lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
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. 555329; 10 µg/mL for coating) and soluble Anti-Human CD28 (Cat. No. 555725; 1 µg/mL) antibodies, and Human Recombinant IL-2 (Cat. No. 554603; 10 ng/mL). Unstimulated PBMC (from the same donor) and the stimulated PBMC were stained with PE Mouse Anti-Human CD8 (Cat. No. 555367/557086/561949/561950), BD Horizon™ BV421 Mouse Anti-Human CD279 (PD-1) (Cat. No. 562516/565935), and BD Horizon™ APC-R700 Mouse Anti-Human LAG-3 (CD223) (Cat. No. 565774/565775) antibodies. Two-color flow cytometric contour 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 unstimulated (Top Plots) or stimulated (Bottom Plots) lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
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
3902
AB_2744329
Aqueous buffered solution containing BSA, protein stabilizer, glycerol 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 APC-R700 under optimum conditions, and unconjugated antibody and free BD Horizon APC-R700 were removed.

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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.

This antibody was conjugated to BD Horizon APC-R700, which has been developed exclusively by BD Biosciences as a better alternative to Alexa Fluor® 700. APC-R700 excites and emits at similar wavelengths to Alexa Fluor® 700 yet exhibits significantly improved brightness. This dye can be excited by the red laser and detected with the same filter set as Alexa Fluor® (eg, 730/45-nm filter).

Format Details
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APC-R700
The BD Horizon™ APC-R700 (APC-R700) Dye is a part of the BD APC red family of dyes. This tandem fluorochrome is comprised of an Allophycocyanin (APC) dye donor that has excitation maximum (Ex Max) of 651-nm and an acceptor dye, R700, with an emission maximum (Em Max) at 706-nm. APC-R700, driven by BD innovation, is designed to be excited by the red (627–640-nm) laser and detected using an optical filter centered near 710-nm (e.g., a 720/40-nm bandpass filter). APC-R700 is a brighter alternative to Alexa Fluor™ 700. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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APC-R700
Red 627-640 nm
651 nm
706 nm
Citations & References
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Development References (5)

  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
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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.