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LAG-3 (human) monoclonal antibody (17B4) (FITC conjugate)

ALX-804-806F-C100 100 µg FITC 486.00 USD
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Product Specification

Alternative Name:Lymphocyte activation gene-3, FDC protein, CD223
Immunogen:Synthetic peptide corresponding to 30 aa (GPPAAAPGHPLAPGPHPAAPSSWGPRPRRY) from the first N-terminal D1 domain of human LAG-3 (lymphocyte activation gene-3).
UniProt ID:P18627
Species reactivity:Human
Specificity:Recognizes the 30 aa extra-loop of the first N-terminal D1 domain of LAG-3.
Applications:Flow Cytometry
Application Notes:Excellent for Flow Cytometry.
Purity Detail:Purified from concentrated hybridoma tissue culture supernatant by Protein A Sepharose™ CL-4B Flow Fast Column.
Formulation:Liquid. In PBS containing 50mM Tris-HCl, 1% BSA and 0.02% sodium azide.
Use/Stability:Stable for at least 6 months after receipt when stored as recommended.
Handling:Do not freeze. Protect from light.
Shipping:Shipped on Blue Ice
Short Term Storage:+4°C
Long Term Storage:+4°C
Scientific Background: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.
Technical Info/Product Notes:SEPHAROSE is a trademark of GE Healthcare companies.
804-806 2
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].
804-806 3
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.
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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, 3x106 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].
804-806 5
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].
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Product Literature References

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;
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
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;
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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