Human LAG-3 Antibody

Catalog # Availability Size / Price Qty
AF2319
AF2319-SP
Detection  of Human LAG‑3 by Western Blot.
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Product Details
Citations (5)
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Human LAG-3 Antibody Summary

Species Reactivity
Human
Specificity
Detects human LAG-3 in direct ELISAs and Western blots. In direct ELISAs, less than 10% cross-reactivity with recombinant mouse LAG-3 is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
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
1 µg/mL
See below
Flow Cytometry
0.25 µg/106 cells
See below
CyTOF-reported
Lowther, D.E. et al. (2016) JCI Insight 1:e85935. 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

Western Blot Detection  of Human LAG‑3 by Western Blot. View Larger

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.

Flow Cytometry Detection of LAG‑3 in CD3+ Human PBMCs by Flow Cytometry. View Larger

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 CD3 epsilon 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.

Flow Cytometry Detection  of LAG-3 in HEK293 Human Cell Line Transfected with Human LAG-3 and eGFP by Flow Cytometry. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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

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.

5 Citations: Showing 1 - 5
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  1. 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;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  2. 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;543(7647):728-732.
    Species: Human
    Sample Types: Tissue Homogenates
    Applications: Functional Assay
  3. Fine needle aspirate flow cytometric phenotyping characterizes immunosuppressive nature of the mesothelioma microenvironment
    Sci Rep, 2016;6(0):31745.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  4. 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;118(4):965-74.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  5. 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, 0;193(6):3101-12.

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