Human LAG-3 Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human LAG-3 in ELISAs.
  • Source
    Monoclonal Mouse IgG1 Clone # 874501
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human LAG-3
    Leu23-Leu450
    Accession # P18627
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    2 µg/mL
    See below
  • Flow Cytometry
    0.25 µg/106 cells
    See below
  • CyTOF-reported
    Cheng, Y. et al. (2016) J. Immunol. 196: 924. Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
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.
Data Examples
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 2 µg/mL of Mouse Anti-Human LAG‑3 Monoclonal Antibody (Catalog # MAB23193) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). 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.
Detection of LAG‑3 in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) were either untreated (top panel) or treated with with 5 μg/mL PHA (bottom panel) for 5 days. PBMCs were stained with Mouse Anti-Human LAG‑3 Monoclonal Antibody (Catalog # MAB23193) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B) and Mouse Anti-Human CD3 epsilon APC‑conjugated Monoclonal Antibody (Catalog # FAB100A). Quadrant markers were set based on isotype control antibody staining (Catalog # MAB002) (data not shown). View our protocol for Staining Membrane-associated Proteins.
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) irrelelvant transfectants and eGFP was stained with Mouse Anti-Human LAG-3 Monoclonal Antibody (Catalog # MAB23193) followed by APC-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). Quadrant markers were set based on control antibody staining (Catalog # MAB002, data not shown). View our protocol for Staining Membrane-associated Proteins.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 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.

  • References:
    1. Triebel, F. et al. (1990) J. Exp. Med. 171:1393.
    2. Baixeras, E. et al. (1992) J. Exp. Med 176:327.
    3. Workman, C.J. and D.A. Vignali (2003) Eur. J. Immunol. 33:970.
    4. Workman, C.J. et al. (2004) J. Immunol. 172:5450.
    5. Huang, C.T. et al. (2004) Immunity 21:503.
    6. Andreae, S. et al. (2003) Blood 102:2130.
  • Long Name:
    Lymphocyte-activation Gene 3
  • Entrez Gene IDs:
    3902 (Human); 16768 (Mouse); 297596 (Rat); 102122272 (Cynomolgus Monkey)
  • Alternate Names:
    CD223 antigen; CD223; FDC; LAG3; LAG-3; lymphocyte activation gene 3 protein; lymphocyte-activation gene 3; Protein FDC
Related Research Areas
Isotype Controls
Description Application Cat# Citations Images  

Mouse IgG1 Isotype Control

Ctrl MAB002 138  
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Mouse IgG1 Isotype Control

Ctrl MAB002R 1
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Secondary Antibodies
Description Application Cat# Citations Images  

Mouse IgG HRP-conjugated Antibody

WB HAF007 26  
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Mouse F(ab)2 IgG (H+L) PE-conjugated Antibody

Flow F0102B 10  
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Donkey Anti-Mouse IgG NorthernLights™ NL557-conjugate Antibody

Flow, IHC, ICC NL007 9
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Donkey Anti-Mouse IgG Biotinylated Antibody

WB BAF018 1
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Mouse IgG HRP-conjugated Antibody

WB HAF018 9  
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Mouse F(ab)2 IgG (H+L) APC-conjugated Antibody

Flow F0101B 2  
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Donkey Anti-Mouse IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL008 8
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Donkey Anti-Mouse IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL009 5
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Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0103B 5  
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Mouse IgG Antibody

WB AF007 4
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Goat Anti-Mouse IgG Biotinylated Antibody

WB BAF007 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Mouse IgG VisUCyte HRP Polymer Antibody

IHC VC001  
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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0114  
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Goat Anti-Mouse IgG Fc Affinity Purified Polyclonal Ab

G-202-C 2
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