Human LAG-3 Antibody (1009611)

R&D Systems | Catalog # MAB23195

R&D Systems
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Human

Applications

Western Blot, Flow Cytometry, CyTOF-ready

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 1009611
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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.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Scientific Data Images for Human LAG-3 Antibody (1009611)

Detection of Human LAG-3 antibody by Western Blot.

Detection of Human LAG‑3 by Western Blot.

Western blot shows lysates of HEK293T human embryonic kidney cell line either mock transfected or transfected with human LAG-3. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human LAG-3 Monoclonal Antibody (Catalog # MAB23195) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for LAG-3 at approximately 100 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of LAG-3 antibody in Human PBMCs antibody by Flow Cytometry.

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

Human peripheral blood mononuclear cells (PBMCs) were either untreated (bottom panel) or treated with with 5 µg/mL PHA (top panel) for 5 days. PBMCs were stained with Mouse Anti-Human LAG-3 Monoclonal Antibody (Catalog # MAB23195) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B) and Mouse Anti-Human CD3 epsilon PE-conjugated Monoclonal Antibody (Catalog # FAB100P). Quadrant markers were set based on isotype control antibody staining (Catalog # MAB003) (data not shown). View our protocol for Staining Membrane-associated Proteins.

Applications for Human LAG-3 Antibody (1009611)

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: Human PBMC treated with PHA

Western Blot

2 µg/mL
Sample: HEK293T human embryonic kidney cell line transfected with human LAG‑3

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.


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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

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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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

Additional LAG-3 Products

Product Documents for Human LAG-3 Antibody (1009611)

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human LAG-3 Antibody (1009611)

For research use only

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FAQs for Human LAG-3 Antibody (1009611)

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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