Human LAG-3 Antibody Summary
Leu23-Leu450
Accession # P18627
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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Detection of Human LAG‑3 by Western Blot. Western blot shows lysates of human peripheral blood mononuclear cells (PBMC) untreated or treated (+) with 1 ug/mL PHA for 5 days and HDLM-2 human Hodgkin's lymphoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human LAG-3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2319) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for LAG-3 at approximately 60-75 kDa (as indicated). GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
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Detection of LAG‑3 in CD3+Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) either (A) untreated or (B) treated with 1 ug/mL PHA for 5 days were stained with Goat Anti-Human LAG-3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2319) followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108) and Mouse Anti-Human CD3e PE-conjugated Monoclonal Antibody (Catalog # FAB100P). Quadrant markers were set based on control antibody staining (Catalog # AB-108-C). View our protocol for Staining Membrane-associated Proteins.
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Detection of LAG-3 in HEK293 Human Cell Line Transfected with Human LAG-3 and eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with either (A) human LAG-3 or (B) irrelevant transfectants and eGFP was stained with Goat Anti-Human LAG-3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2319) followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). Quadrant markers were set based on control antibody staining(Catalog # AB-108-C, data not shown). View our protocol for Staining Membrane-associated Proteins.
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LAG‑3 in Human Spleen. LAG‑3 was detected in immersion fixed paraffin-embedded sections of human spleen using Goat Anti-Human LAG‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2319) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
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LAG‑3 in Mouse Spleen. LAG‑3 was detected in immersion fixed paraffin-embedded sections of mouse spleen using Goat Anti-Human LAG‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2319) at 15 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Reconstitution Calculator
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: LAG-3
LAG-3 (Lymphocyte activation gene-3), also known as CD223, is a member of the immunoglobulin superfamily (IgSF). The mature LAG-3 protein is a 496 amino acid (aa) membrane protein with a 421 aa extracellular region which contains four IgSF domains, a 21 aa transmembrane region and a 54 aa cytoplasmic region. LAG-3 and CD4 molecules share < 20% aa sequence homology but have a similar structure (1, 2). Both molecules bind to MHC class II. LAG-3 binds to MHC class II with higher affinity compared to CD4. Both LAG-3 and CD4 genes are located on the distal part of the short arm of chromosome 12. LAG-3 is an activation-induced molecule, expressed on activated T cells and NK cells, but not on resting T cells. Studies using LAG-3 -/- mice have shown significant delay of T cell apoptosis following antigen stimulation and increased size of memory T cells pool following infection (3, 4). It also has been reported that anti-LAG-3 antibodies up-regulate T cell activation by blocking interaction of LAG-3 and MHC class II. The study has demonstrated that LAG-3 is selectively expressed on activated CD4+CD25+ TReg cells and plays a role in their suppressive activity (5). This evidence indicated, unlike the interaction of CD4 with MHC class II that plays a positive role in T cell activation, LAG-3 binds to MHC class II and negatively regulates T cell activation through LAG-3 signaling. On the other hand, studies have shown that binding of LAG-3 to MHC class II molecules on antigen presenting cells induce maturation of dendritic cells and cytokine secretion by monocytes through MHC class II signal transduction (6). Taken together, LAG-3 may have two major functions, it negatively regulates T cells activation through LAG-3 signaling and stimulates antigen presenting cells which express MHC class II.
- Triebel, F. et al. (1990) J. Exp. Med. 171:1393.
- Baixeras, E. et al. (1992) J. Exp. Med 176:327.
- Workman, C.J. and D.A. Vignali (2003) Eur. J. Immunol. 33:970.
- Workman, C.J. et al. (2004) J. Immunol. 172:5450.
- Huang, C.T. et al. (2004) Immunity 21:503.
- Andreae, S. et al. (2003) Blood 102:2130.
Product Datasheets
Citations for Human LAG-3 Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 5
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Low and variable tumor reactivity of the intratumoral TCR repertoire in human cancers
Authors: W Scheper, S Kelderman, LF Fanchi, C Linnemann, G Bendle, MAJ de Rooij, C Hirt, R Mezzadra, M Slagter, K Dijkstra, RJC Kluin, P Snaebjorns, K Milne, BH Nelson, H Zijlmans, G Kenter, EE Voest, JBAG Haanen, TN Schumacher
Nat. Med., 2018-12-03;0(0):.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry -
Effective combinatorial immunotherapy for castration-resistant prostate cancer
Authors: X Lu, JW Horner, E Paul, X Shang, P Troncoso, P Deng, S Jiang, Q Chang, DJ Spring, P Sharma, JA Zebala, DY Maeda, YA Wang, RA DePinho
Nature, 2017-03-20;543(7647):728-732.
Species: Human
Sample Types: Tissue Homogenates
Applications: Functional Assay -
Fine needle aspirate flow cytometric phenotyping characterizes immunosuppressive nature of the mesothelioma microenvironment
Sci Rep, 2016-08-19;6(0):31745.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry -
Trafficking of LAG-3 to the surface on activated T cells via its cytoplasmic domain and protein kinase C signaling.
Authors: Bae J, Lee S, Park C, Lee Y, Chun T
J Immunol, 2014-08-08;193(6):3101-12.
Species: Human
Sample Types: Cell Lysates, Whole Cells
Applications: Functional Assay, ICC, Western Blot -
Responsiveness of HIV-specific CD4 T cells to PD-1 blockade.
Authors: Porichis F, Kwon DS, Zupkosky J, Tighe DP, McMullen A, Brockman MA, Pavlik DF, Rodriguez-Garcia M, Pereyra F, Freeman GJ, Kavanagh DG, Kaufmann DE
Blood, 2011-06-07;118(4):965-74.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry
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