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 Alexa Fluor® 647 Mouse Anti-Human LAG-3 (CD223) (Cat. No. 565716/565717) 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 Alexa Fluor® 647 Mouse Anti-Human LAG-3 (CD223) (Cat. No. 565716/565717) 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
BD Pharmingen™
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_2744328
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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
Product Notices
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO
Size
50 µg
Cat No.
565571
Purified NA/LE Mouse Anti-Human CD3 RUO
Size
0.5 mg
Cat No.
555329
Purified NA/LE Mouse Anti-Human CD28 RUO
Size
0.5 mg
Cat No.
555725
Recombinant Human IL-2 RUO
Size
10 µg
Cat No.
554603
565716 Rev. 1
Antibody Details
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.
565716 Rev. 1
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
565716 Rev.1
Citations & References
View product citations for antibody "565716" on CiteAb
Development References (5)
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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
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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
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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
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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
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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
565716 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.