The lymphocyte activation gene-3 (LAG-3, CD223), a member of the immunoglobulin superfamily (IgSF) related to CD4, binds to the major histocompatibility complex (MHC) class II molecules but with higher affinity than CD4. Several alternative mRNA splice-variants of human LAG-3 have been described, two of them encoding potential secreted forms: LAG-3V1 (i.e. the D1-D2 domains of the protein, 36 kDa) and LAG-3V3 (D1-D3, 52 kDa). The longer form was detected by ELISA in the serum of healthy individuals as well as of tuberculosis patients with a favorable outcome. LAG-3 expression by T cell clones correlated with IFN-γ production, and hence soluble LAG-3 has been suggested as a serological marker of Th1 responses.
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Figure: LAG-3 expression on activated human peripheral blood mononuclear cells (PBMC) detected with LAG-3 (human), mAb (17B4) (FITC) (Prod. No. ALX-804-806F).Method: T lymphocytes from human PBMC are stimulated with 1µg/ml of PHA for three days. Then, after seven days of culture, 3×106 three-days PHA-activated human PBMC are treated with LAG-3 (human), mAb (17B4) (FITC) (Prod. No. ALX-804-806F) or FITC coupled isotype-matched (IgG1) control MAb (used at a saturating dilution of 1/800 and 1/150 respectively) for 30 min. at 4°C in RPMI 1640 and washed twice with 1x PBS. Stained cells are then analysed by FACS [4].

Figure: Expression of LAG-3 on CD4+ and CD8+ subpopulations of tumour infiltrating lymphocytes (TILs) detected with LAG-3 (human), mAb (17B4) (FITC) (Prod. No. ALX-804-806F).Method: TILs from a dissociated renal cell carcinoma sample, stained with 5µg/ml LAG-3 (human), mAb (17B4) (FITC) (Prod. No. ALX-804-806F) and FITC-coupled anti-CD4 or -CD8, are analyzed by a two-colour FACS analysis. Additional staining with anti-CD3 allowed a gate analysis of total T cells. Values indicate percentages in each quadrant [5].

Figure: Specific inhibition of 17B4 staining.Method: LAG-3 (human), mAb (17B4) (FITC) (Prod. No. ALX-804-806F) (10µg/ml) (is preincubated with a specific peptide epitope (208b) or a control tetanus toxoid (TT) peptide at different molarities prior to staining of TILs. Stained cells are then analyzed by FACS.

Figure: Tumor infiltrating lymphocytes (TILs) express LAG-3 (detected using LAG-3 (human), mAb (17B4) (Prod. No. ALX-804-806)). Method: Freshly dissociated single cell suspensions of renal cell carcinoma TILs are incubated with LAG-3 (human), mAb (17B4) (FITC) (Prod. No. ALX-804-806) (5µg/ml) and anti-MHC Class II molecules (PE) for 30 min. and washed twice in saline buffer. Additional staining with anti-CD3 allowed a gate analysis of total T cells. The LAG-3 and MHC II profiles of CD3+-gated cells for 3 patients are shown [5].




Product Details
Alternative Name |
Lymphocyte activation gene-3, FDC protein, CD223 |
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Application |
Flow Cytometry |
Application Notes |
Excellent for Flow Cytometry. |
Clone |
17B4 |
Formulation |
Liquid. In PBS containing 50mM Tris-HCl, 1% BSA and 0.02% sodium azide. |
Host |
Mouse |
Immunogen |
Synthetic peptide corresponding to 30 aa (GPPAAAPGHPLAPGPHPAAPSSWGPRPRRY) from the first N-terminal D1 domain of human LAG-3 (lymphocyte activation gene-3). |
Isotype |
IgG1 |
Purity Detail |
Purified from concentrated hybridoma tissue culture supernatant by Protein A Sepharose™ CL-4B Flow Fast Column. |
Source |
Purified from hybridoma tissue culture supernatant. |
Species Reactivity |
Human |
Specificity |
Recognizes the 30 aa extra-loop of the first N-terminal D1 domain of LAG-3. |
Technical Info / Product Notes |
SEPHAROSE is a trademark of GE Healthcare companies. |
UniProt ID |
P18627 |
Worry-free Guarantee |
This antibody is covered by our Worry-Free Guarantee. |
Handling & Storage
Use/Stability |
Stable for at least 6 months after receipt when stored as recommended. |
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Handling |
Do not freeze. Protect from light. |
Short Term Storage |
+4°C |
Long Term Storage |
+4°C |
Shipping |
Blue Ice |
Regulatory Status |
RUO – Research Use Only |
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- Preoperative immune checkpoint inhibition and cryoablation in early-stage breast cancer: Comen, E., Budhu, S., et al.; iScience 27, 108880 (2024), Abstract
- Nivolumab plus ipilimumab in advanced salivary gland cancer: a phase 2 trial: Vos, J., Burman, B., et al.; Nat. Med. 29, 3077 (2023), Abstract
- Brain radiotherapy, tremelimumab-mediated CTLA-4-directed blockade +/− trastuzumab in patients with breast cancer brain metastases: D.B. Page, et al.; NPJ Breast Cancer 8, 50 (2022), Abstract
- Immunological status of peripheral blood is associated with prognosis in patients with bone and soft-tissue sarcoma: Kim, Y., Kobayashi, E., et al.; Oncol. Lett. 21, 212 (2021), Abstract
- Analysis of CAR-Mediated Tonic Signaling: H. Calderon, et al.; Methods Mol. Biol. 2086, 223 (2020), Abstract
- Targeting VEGFR2 with Ramucirumab strongly impacts effector/ activated regulatory T cells and CD8+ T cells in the tumor microenvironment: Tada, Y., Togashi, Y., et al.; J. Immunother. Cancer 6, 106 (2018), Abstract
- Improving CART-Cell Therapy of Solid Tumors with Oncolytic Virus-Driven Production of a Bispecific T-cell Engager: A. Wing, et al.; Cancer Immunol. Res. 6, 605 (2018), Application(s): Flow Cytometry, Abstract
- NK cell and IFN signatures are positive prognostic biomarkers for resectable pancreatic cancer: M. Hoshikawa, et al.; Biochem. Biophys. Res. Commun. 495, 2058 (2018), Application(s): Flow cytometry, Abstract
- PEGylated IL-10 (Pegilodecakin) Induces Systemic Immune Activation, CD8+ T Cell Invigoration and Polyclonal T Cell Expansion in Cancer Patients: A. Naing, et al.; Cancer Cell 34, 775 (2018), Abstract
- Tumor-infiltrating and peripheral blood T cell immunophenotypes predict early relapse in localized clear cell renal cell carcinoma: N.A. Giraldo, et al.; Clin. Cancer Res. 23, 4416 (2017), Application(s): Flow cytometry, Abstract
- CD26-Mediated Induction of EGR2 and IL-10 as Potential Regulatory Mechanism for CD26 Costimulatory Pathway: R. Hatano, et al.; J. Immunol. 194, 960 (2015), Abstract
- Elevated expression of LAG-3, but not PD-1, is associated with impaired iNKT cytokine production during chronic HIV-1 infection and treatment: J.A. Juno, et al.; Retrovirology 12, 17 (2015), Application(s): Flow Cytometry, Abstract — Full Text
- Enrichment of LAG-3, but not PD-1, on Double Negative T Cells at the Female Genital Tract: J.A. Juno, et al.; Am. J. Reprod. Immunol. 72, 534 (2014), Abstract
- Fusion of Ubiquitin to HIV Gag Impairs Human Monocyte-Derived Dendritic Cell Maturation and Reduces Ability to Induce Gag T Cell Responses: S. Herath, et al.; PLoS One 9, e88327 (2014), Application(s): Flow cytometry using human T-cells, Abstract — Full Text
- PD-1 identifies the patient-specific CD8+ tumor-reactive repertoire infiltrating human tumors: A. Gros, et al.; J. Clin. Invest. 124, 2246 (2014), Application(s): Flow cytometry using human peripheral blood lymphocytes (PBL) and metastatic melanoma cells, Abstract — Full Text
- Selection of CD8+PD-1+ lymphocytes in fresh human melanomas enriches for tumor-reactive T cells: T. Inozume, et al.; J. Immunother. 33, 956 (2010), Application(s): FC analysis with human tumor digests or cultured tumor-infiltrating lymphocytes, Abstract — Full Text
- Human lymphocyte activation gene-3 molecules expressed by activated T cells deliver costimulation signal for dendritic cell activation: C. Casati, et al.; J. Immunol. 180, 3782 (2008), Abstract
- Human dendritic cells acquire a semimature phenotype and lymph node homing potential through interaction with CD4+CD25+ regulatory T cells: J.Bayry, et al.; J. Immunol. 178, 4184 (2007), Application(s): FC analysis with human regulatory T cells, Abstract — Full Text
- A soluble lymphocyte activation gene-3 (sLAG-3) protein as a prognostic factor in human breast cancer expressing estrogen or progesterone receptors: F. Triebel, et al.; Cancer Lett. 235, 147 (2006), Abstract
- Immunological mechanisms elicited at the tumour site by lymphocyte activation gene-3 (LAG-3) versus IL-12: sharing a common Th1 anti-tumour immune pathway: E.D. Carlo, et al.; J. Pathol. 205, 82 (2005), Abstract
- The negative regulatory function of the lymphocyte-activation gene-3 co-receptor (CD223) on human T cells: L. Macon-Lemaitre and F. Triebel; Immunology 115, 170 (2005), Abstract
- Responses to human CD40 ligand/human interleukin-2 autologous cell vaccine in patients with B-cell chronic lymphocytic leukemia: E. Biagi, et al.; Clin. Cancer Res. 11, 6916 (2005), Abstract — Full Text
- T Lymphocytes infiltrating various tumour types express the MHC class II ligand lymphocyte activation gene-3 (LAG-3): role of LAG-3/MHC class II interactions in cell-cell contacts: C.E. Demeure, et al.; Eur. J. Cancer 37, 1709 (2001), Abstract
- CD3/TCR complex-associated lymphocyte activation gene-3 molecules inhibit CD3/TCR signaling: S. Hannier, et al.; J. Immunol. 161, 4058 (1998), Abstract — Full Text
- Characterization of the major histocompatibility complex class II binding site on LAG-3 protein: B. Huard, et al.; PNAS 94, 5744 (1997), Abstract — Full Text
- Cellular expression and tissue distribution of the human LAG-3-encoded protein, an MHC class II ligand: B. Huard, et al.; Immunogenetics 39, 213 (1994), Abstract
- Characterization of the lymphocyte activation gene 3-encoded protein. A new ligand for human leukocyte antigen class II antigens: E. Baixeras, et al.; J. Exp. Med. 176, 327 (1992), Abstract — Full Text
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