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Anti-LAG-3 antibody [EPR20294-77] (ab209238) is a rabbit monoclonal antibody that is used to detect LAG-3 in Western Blot, IP, ICC/IF. Suitable for Mouse samples.



- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency


Images

Immunocytochemistry/ Immunofluorescence - Anti-LAG-3 antibody [EPR20294-77] (AB209238), expandable thumbnail
  • Western blot - Anti-LAG-3 antibody [EPR20294-77] (AB209238), expandable thumbnail
  • Immunoprecipitation - Anti-LAG-3 antibody [EPR20294-77] (AB209238), expandable thumbnail
  • Western blot - Anti-LAG-3 antibody [EPR20294-77] (AB209238), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBICC/IF
Mouse
Tested
Tested
Tested

Tested
Tested

Species
Mouse
Dilution info
1/30
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/1000
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/100
Notes

-

Associated Products

Select an associated product type

15 products for Alternative Product

Target data

Function

Lymphocyte activation gene 3 protein: Inhibitory receptor on antigen activated T-cells (PubMed:12209638, PubMed:12421911, PubMed:12672063, PubMed:15100286, PubMed:15634887, PubMed:30580966). Delivers inhibitory signals upon binding to ligands, such as FGL1 (PubMed:30580966). FGL1 constitutes a major ligand of LAG3 and is responsible for LAG3 T-cell inhibitory function (PubMed:30580966). Following TCR engagement, LAG3 associates with CD3-TCR in the immunological synapse and directly inhibits T-cell activation (PubMed:12209638, PubMed:12421911, PubMed:12672063, PubMed:15100286, PubMed:15634887). May inhibit antigen-specific T-cell activation in synergy with PDCD1/PD-1, possibly by acting as a coreceptor for PDCD1/PD-1 (PubMed:21300912). Negatively regulates the proliferation, activation, effector function and homeostasis of both CD8(+) and CD4(+) T-cells (PubMed:12209638, PubMed:12421911, PubMed:12672063, PubMed:15100286, PubMed:15634887). Also mediates immune tolerance: constitutively expressed on a subset of regulatory T-cells (Tregs) and contributes to their suppressive function (PubMed:15485628). Also acts as a negative regulator of plasmacytoid dendritic cell (pDCs) activation (PubMed:19201841). Binds MHC class II (MHC-II); the precise role of MHC-II-binding is however unclear (PubMed:12209638, PubMed:12421911, PubMed:15634887). Secreted lymphocyte activation gene 3 protein. May function as a ligand for MHC class II (MHC-II) on antigen-presenting cells (APC), promoting APC activation/maturation and driving Th1 immune response.

Alternative names

CD223, Lymphocyte activation gene 3 protein, LAG-3, Activation-induced cytidine deaminase-linked autoimmunity protein, Aida, Lag3

Recommended products

Anti-LAG-3 antibody [EPR20294-77] (ab209238) is a rabbit monoclonal antibody that is used to detect LAG-3 in Western Blot, IP, ICC/IF. Suitable for Mouse samples.



- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR20294-77
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

What is this antibody validated in?


Anti-LAG-3 antibody [EPR20294-77] (ab209238) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Immunoprecipitation (IP), Immunocytochemistry/immunofluorescence (ICC/IF), in Mouse samples.

What is the molecular weight of LAG-3?


Anti-LAG-3 [EPR20294-77] (ab209238) specifically detects a band for LAG-3 (UniProt: Q61790) at a molecular weight of 57kDa.

Recommended positive controls


WB: Mouse LAG-3 recombinant protein (aa24-442) tagged with murine IgG2A at the C-terminus; mouse splenocytes treated with 5 ug/ml concanavalin A for 72 hours, whole cell lysate.ICC/IF: Mouse splenocytes.IP: Mouse splenocytes treated with 5 µg/ml concanavalin A for 72 hours, whole cell lysate.

Trial sizes available!


Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.Other related products


We have a range of other formats of antibody clone [EPR20294-77] also available for your convenience:
ab209238, Carrier free - Anti-LAG-3 antibody [EPR20294-77] - BSA and Azide free ab236154



Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

LAG-3 also known as CD223 is a transmembrane protein with a mass of approximately 70 kDa. It acts as a checkpoint molecule similar to CTLA-4 and PD-1 in the immune system. LAG-3 is expressed on activated T cells natural killer cells and some dendritic cells. It interacts with its ligand MHC class II leading to inhibition of cellular proliferation and cytokine secretion an important part of immune response regulation.

Biological function summary

LAG-3 is involved in maintaining immune homeostasis and preventing autoimmunity. It is not part of a complex but works in synergy with other immune checkpoint molecules. When interacting with MHC class II LAG-3 transmits inhibitory signals that attenuate immune cell responses. Its role in immune regulation helps to balance immune activation and tolerance which is essential for preventing tissue damage while protecting against pathogens.

Pathways

The inhibitory role of LAG-3 integrates into the T cell receptor (TCR) signaling pathway and is also linked to the PD-1 signaling pathway. In TCR signaling LAG-3 negatively regulates T cell activation. LAG-3 is functionally related to proteins like PD-1 and CTLA-4 sharing similar roles in immune checkpoint pathways that are critical for maintaining immune balance.

Associated diseases and disorders

LAG-3 has links to cancer and autoimmune diseases. In cancer LAG-3 expression on tumor-infiltrating lymphocytes correlates with immune evasion by tumors. This relationship appears in various cancers including melanoma and renal cell carcinoma. In autoimmune diseases LAG-3's expression on regulatory T cells helps control conditions such as multiple sclerosis. It collaborates with CTLA-4 in modulating immune responses that prevent excessive immune activation and tissue damage associated with autoimmunity.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

4 product images

  • Immunocytochemistry/ Immunofluorescence - Anti-LAG-3 antibody [EPR20294-77] (ab209238), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-LAG-3 antibody [EPR20294-77] (ab209238)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized untreated and concanavalin A (2.5 μg/ml for 72 hours) treated mouse splenocytes labeling LAG-3 with ab209238 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing increased membranous staining following treatment of mouse splenocytes with concanavalin A (2.5 μg/ml for 72 hours) (PMID 20391435; PMID 26318293; PMID 11527700; PMID 19631993).

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)(Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution

  • Western blot - Anti-LAG-3 antibody [EPR20294-77] (ab209238), expandable thumbnail

    Western blot - Anti-LAG-3 antibody [EPR20294-77] (ab209238)

    Blocking/Dilution Buffer: 5% NFDM/TBST

    All lanes: Western blot - Anti-LAG-3 antibody [EPR20294-77] (ab209238) at 1/10000 dilution

    All lanes: Mouse LAG-3 recombinant protein (aa24-442) tagged with murine IgG2A at the C-terminus, 10 ng

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 57 kDa

    Observed band size: 100 kDa

    Exposure time: 1s

  • Immunoprecipitation - Anti-LAG-3 antibody [EPR20294-77] (ab209238), expandable thumbnail

    Immunoprecipitation - Anti-LAG-3 antibody [EPR20294-77] (ab209238)

    LAG-3 was immunoprecipitated from 0.35 mg of mouse splenocytes (treated with 5 μg/ml concanavalin A for 72 hours) whole cell lysate with ab209238 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab209238 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/1000 dilution.

    Lane 1: Mouse splenocytes (treated with 5 μg/ml concanavalin A for 72 hours), whole cell lysate 10μg (Input).

    Lane 2: ab209238 IP in mouse splenocytes (treated with 5 μg/ml concanavalin A for 72 hours), whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab209238 in mouse splenocytes (treated with 5 μg/ml concanavalin A for 72 hours), whole cell lysate.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 1 second.

    All lanes: Immunoprecipitation - Anti-LAG-3 antibody [EPR20294-77] (ab209238)

    Predicted band size: 57 kDa

    Observed band size: 54 kDa, 70 kDa

  • Western blot - Anti-LAG-3 antibody [EPR20294-77] (ab209238), expandable thumbnail

    Western blot - Anti-LAG-3 antibody [EPR20294-77] (ab209238)

    Blocking/Dilution Buffer: 5% NFDM/TBST

    The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

    LAG3 expression can be elevated in activated splenocytes (PMID 19631993).

    The molecular mass observed is consistent with the literature (PMID 15971272; PMID 15557174).

    All lanes: Western blot - Anti-LAG-3 antibody [EPR20294-77] (ab209238) at 1/1000 dilution

    Lane 1: Untreated mouse splenocytes whole cell lysate at 10 µg

    Lane 2: Mouse splenocytes treated with 5 ug/ml concanavalin A for 72 hours whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 57 kDa

    Observed band size: 54 kDa, 70 kDa

    Exposure time: 10s

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Product protocols

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