Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Primate - Macaca fascicularis (Crab-eating Monkey or Cynomolgus Macaque), Primate - Macaca mulatta (Rhesus Macaque), Xenograft

Applications

Validated:

Flow Cytometry

Cited:

Flow Cytometry, Bioassay, CyTOF-reported

Label

Phycoerythrin (Excitation = 488 nm, Emission = 565-605 nm)

Antibody Source

Polyclonal Goat IgG
View all LAG-3 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human LAG-3
Leu23-Leu450
Accession # P18627

Specificity

Detects human LAG-3 in direct ELISAs and Western blots. In direct ELISAs and Western blots, less than 1% cross-reactivity with recombinant mouse LAG-3 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human LAG‑3 PE‑conjugated Antibody

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

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 Phycoerythrin-conjugated Antigen Affinity-purified Polyclonal Antibody (Catalog # FAB2319P). Quadrant markers were set based on control antibody staining (Catalog # IC108P, data not shown). View our protocol for Staining Membrane-associated Proteins.
Detection of LAG-3 antibody in Human PBMCs antibody by Flow Cytometry.

Detection of LAG-3 in Human PBMCs by Flow Cytometry.

CD3+ PHA-treated peripheral blood mononuclear cells (PBMCs) were stained with Mouse Anti-Human CD3e APC-conjugated Monoclonal Antibody (Catalog # FAB100A) and either (A) Goat Anti-Human LAG-3 PE-conjugated Antigen Affinity-purified Polyclonal Antibody (Catalog # FAB2319P) or (B) Normal Goat IgG Phycoerythrin Control (Catalog # IC108P). View our protocol for Staining Membrane-associated Proteins.
Detection of Human LAG-3 by Flow Cytometry

Detection of Human LAG-3 by Flow Cytometry

Human CD4+CD25−LAG3+ T cells suppressed antibody production. a Gating strategy of fluorescence-activated cell sorting analysis for CD4+CD25−LAG3+CD45RA− T cells (LAG3+ Tregs) and CD4+CD25+CD127dimCD45RA− T cells (CD25+ Tregs). b IL-10, IFN-gamma, and IL-17A protein levels in the culture supernatants after 3 days of incubation of the indicated T-cell subsets was determined by ELISA (n = 6). c B cells and TFH cells were cocultured with the indicated subsets for 12 days in the presence of SEB stimulation, and total IgG, IgM, and IgA production was determined by ELISA (n = 6). d B cells and TFH cells were cocultured with CD4+CD25−LAG3+CD49b− T cells for 12 days in the presence of SEB stimulation, and total IgG was determined by ELISA (n = 3). All error bars represent SD. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. All P values were calculated by one-way analysis of variance and Tukey’s multiple-comparisons test. CD Cluster of differentiation, ELISA Enzyme-linked immunosorbent assay, IFN-gamma Interferon-gamma, Ig Immunoglobulin, IL Interleukin, LAG3 Lymphocyte activation gene 3, RA Rheumatoid arthritis, SEB Staphylococcal enterotoxin B, TFH Follicular helper T cell, Treg Regulatory T cell Image collected and cropped by CiteAb from the following publication (https://arthritis-research.biomedcentral.com/articles/10.1186/s13075-01…), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human LAG-3 by Flow Cytometry

Detection of Human LAG-3 by Flow Cytometry

Effect of abatacept on the CD4+CD25−LAG3+ T-cell subset in vitro. a Naive CD4+ T cells and antigen-presenting cells (APCs) taken from healthy donors were cultured with anti-CD3 antibody in the presence of abatacept, and FACS was conducted on day 4. Representative FACS data from three independent experiments are shown. Percentages of CD4+CD25−LAG3+ T cells in CD4+ T cells are presented in the bar graph (n = 3). b Quantitative real-time polymerase chain reaction analysis of EGR2, FAS, and CD274 (PDL1) mRNA expression in the cells that were induced from naive CD4+ T cells in the presence of abatacept (n = 3). c Sorted 7-amino actinomycin D-negative CD4+ T cells were cultured with B cells and TFH cells under staphylococcal enterotoxin B stimulation for 12 days, and total IgG production was determined by ELISA (n = 3). All error bars represent SD. ** P < 0.001; **** P < 0.0001. All P values were calculated by unpaired two-tailed Student’s t test. CD Cluster of differentiation 4, EGR2 Early growth response gene 2, FACS Fluorescence-activated cell sorting, GAPDH Glyceraldehyde 3-phosphate dehydrogenase, IgG Immunoglobulin G, LAG3 Lymphocyte activation gene 3, mRNA Messenger RNA, PDL1 Programmed death-ligand 1, TFH Follicular helper T cell Image collected and cropped by CiteAb from the following publication (https://arthritis-research.biomedcentral.com/articles/10.1186/s13075-01…), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human LAG‑3 PE‑conjugated Antibody

Application
Recommended Usage

Flow Cytometry

10 µL/106 cells
Sample: Human peripheral blood mononuclear cells (PBMCs) treated with PHA and HEK293 Human Cell Line Transfected with Human LAG-3 and eGFP

Spectra Viewer

Plan Your Experiments

Use our spectra viewer to interactively plan your experiments, assessing multiplexing options. View the excitation and emission spectra for our fluorescent dye range and other commonly used dyes.

Spectra Viewer
Spectra Viewer

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Formulation

Supplied in a saline solution containing BSA and Sodium Azide.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Protect from light. Do not freeze.
  • 12 months from date of receipt, 2 to 8 °C as supplied.

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

Alternate Names

CD223, LAG3

Entrez Gene IDs

3902 (Human); 16768 (Mouse); 297596 (Rat); 102122272 (Cynomolgus Monkey)

Gene Symbol

LAG3

UniProt

P18627

Additional LAG-3 Products

Product Documents for Human LAG‑3 PE‑conjugated Antibody

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Product Specific Notices for Human LAG‑3 PE‑conjugated Antibody

For research use only

Citations for Human LAG‑3 PE‑conjugated Antibody

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FAQs for Human LAG‑3 PE‑conjugated Antibody

Showing  1 - 2 of 2 FAQs Showing All

  • Q: Can this LAG-3 antibody be used in ELISA? 

    A: We have not validated this particular LAG-3 antibody for ELISA in mouse; however, it has been cited as being used as a detection antibody in a publicatoin (PMID 15946792).

  • Q: I'm using your LAG-3 antibody in western blot but the observed band is a different size than the predicted band size, do you know what the bands are?

    A: There are several alternative splice varients of LAG-3 that could explain the different sized proteins stained by the LAG-3 antibody. It is also believed that there are two secreted forms of LAG-3, one that is about 36 kDa and another at 52 kDa.

  • Q: Can this LAG-3 antibody be used in ELISA? 

    A: We have not validated this particular LAG-3 antibody for ELISA in mouse; however, it has been cited as being used as a detection antibody in a publicatoin (PMID 15946792).

  • Q: I'm using your LAG-3 antibody in western blot but the observed band is a different size than the predicted band size, do you know what the bands are?

    A: There are several alternative splice varients of LAG-3 that could explain the different sized proteins stained by the LAG-3 antibody. It is also believed that there are two secreted forms of LAG-3, one that is about 36 kDa and another at 52 kDa.

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