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Rabbit Recombinant Monoclonal LAG-3 antibody. Carrier free. Suitable for ICC/IF, IP, Flow Cyt, WB, IHC-P and reacts with Transfected cell line, Human samples.

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Images

Flow Cytometry - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (AB282638), expandable thumbnail
  • Western blot - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (AB282638), expandable thumbnail
  • Flow Cytometry - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (AB282638), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (AB282638), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (AB282638), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Constituents: 100% PBS

Form
Liquid
Clonality
Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ICC/IFIPFlow CytWBIHC-P
Human
Expected
Expected
Tested
Tested
Tested
Transfected cell line
Tested
Not recommended
Tested
Not recommended
Not recommended

Tested
Tested

Species
Transfected cell line
Dilution info
-
Notes

-

Expected
Expected

Species
Human
Dilution info
Use at an assay dependent concentration.
Notes

-

Expected
Expected

Species
Human
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Transfected cell line
Dilution info
-
Notes

-

Tested
Tested

Species
Transfected cell line, Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Transfected cell line
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

Different mRNA expression levels of LAG3 in brain have been reported in the literature (PMID: 1692078; PMID: 12825348). In IHC, under our experimental conditions, this antibody showed no positive staining on human cerebral cortex.

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species
Transfected cell line
Dilution info
-
Notes

-

Associated Products

Select an associated product type

15 products for Alternative Product

6 products for Alternative Version

Target data

Function

Lymphocyte activation gene 3 protein: Inhibitory receptor on antigen activated T-cells (PubMed:20421648, PubMed:7805750, PubMed:8647185). Delivers inhibitory signals upon binding to ligands, such as FGL1 (By similarity). FGL1 constitutes a major ligand of LAG3 and is responsible for LAG3 T-cell inhibitory function (By similarity). Following TCR engagement, LAG3 associates with CD3-TCR in the immunological synapse and directly inhibits T-cell activation (By similarity). May inhibit antigen-specific T-cell activation in synergy with PDCD1/PD-1, possibly by acting as a coreceptor for PDCD1/PD-1 (By similarity). Negatively regulates the proliferation, activation, effector function and homeostasis of both CD8(+) and CD4(+) T-cells (PubMed:20421648, PubMed:7805750, PubMed:8647185). Also mediates immune tolerance: constitutively expressed on a subset of regulatory T-cells (Tregs) and contributes to their suppressive function (By similarity). Also acts as a negative regulator of plasmacytoid dendritic cell (pDCs) activation (By similarity). Binds MHC class II (MHC-II); the precise role of MHC-II-binding is however unclear (PubMed:8647185). 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, FDC, LAG3, Lymphocyte activation gene 3 protein, LAG-3

Recommended products

Rabbit Recombinant Monoclonal LAG-3 antibody. Carrier free. Suitable for ICC/IF, IP, Flow Cyt, WB, IHC-P and reacts with Transfected cell line, Human samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free
Yes
Clone number
EPR20261
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze

Notes

ab282638 is a low endotoxin version of Anti-LAG-3 antibody [EPR20261] - BSA and Azide free ab227579.

Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

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

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.

7 product images

  • Flow Cytometry - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (ab282638), expandable thumbnail

    Flow Cytometry - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (ab282638)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-LAG-3 antibody [EPR20261] ab209236).

    Flow cytometric analysis of Human peripheral blood mononuclear cells treated with 1 μg/mL PHA for 3 days cells with Anti-LAG-3 antibody [EPR20261] ab209236 at 1/50 dilution (right) compared with a rabbit monoclonal IgG isotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730; left). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 at 1/2000 dilution was used as the secondary antibody.

    Only the CD3+ population are also positive for LAG-3. Gated on total viable cells.

  • Western blot - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (ab282638), expandable thumbnail

    Western blot - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (ab282638)

    This IHC data was generated using the same anti-LAG-3 antibody clone, EPR20261, in a different buffer formulation (Anti-LAG-3 antibody [EPR20261] ab209236).

    Primary loading control and concentration: Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) at 1/20000 dilution

    Secondary loading control and concentration: Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/10000 dilution

    Lanes 1-2: Merged signal (red and green). Green – Anti-LAG-3 antibody [EPR20261] ab209236 observed at 54-70 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
    Anti-LAG-3 antibody [EPR20261] ab209236 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800RCW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

    The expression profile observed in Jurkat is consistent with the literature (PMID: 25108024).

    Negative control: Jurkat (PMID: 25108024)

    Observed MW: 54-70 kDa

    All lanes: Western blot - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (ab282638) at 1/500 dilution

    Lane 1: HDLM-2 (Human Hodgkin lymphoma ) whole cell lysate at 20 µg

    Lane 2: Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution

    Predicted band size: 57 kDa

  • Flow Cytometry - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (ab282638), expandable thumbnail

    Flow Cytometry - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (ab282638)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-LAG-3 antibody [EPR20261] ab209236).

    Flow cytometric analysis of HEK-293T (Human epithelial cell line from embryonic kidney) cells transfected with a GFP-tagged human LAG3 construct labeling LAG-3 with Anti-LAG-3 antibody [EPR20261] ab209236 at 1/500 dilution (right) compared with a rabbit monoclonal IgG isotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730; left). Goat anti rabbit IgG (Alexa Fluor® 647) Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) ab150079 at 1/2000 dilution was used as the secondary antibody.

    Note: Fresh cells without fixation and permeabilization were used to perform FC testing. Only GFP positive population results in LAG3 positive staining (Q2, right panel).

  • Immunocytochemistry/ Immunofluorescence - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (ab282638), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (ab282638)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-LAG-3 antibody [EPR20261] ab209236).

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (Human epithelial cell line from embryonic kidney) cells transfected with GFP-tagged LAG3 expression construct or GFP only, labeling LAG-3 with Anti-LAG-3 antibody [EPR20261] ab209236 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 647) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) ab150079) secondary antibody at 1/1000 dilution (green).

    Confocal image showing positive staining on HEK-293T cells transfected with a GFP-tagged LAG-3 expression construct.

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

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 647) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) ab150079) at 1/1000 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (ab282638), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (ab282638)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-LAG-3 antibody [EPR20261] ab209236).

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling LAG-3 with Anti-LAG-3 antibody [EPR20261] ab209236 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Sporadic cytoplasmic staining on immunocytes of human tonsil [PMID: 11527700; PMID: 16757686].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (ab282638), expandable thumbnail

    Western blot - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (ab282638)

    This IHC data was generated using the same anti-LAG-3 antibody clone, EPR20261, in a different buffer formulation (Anti-LAG-3 antibody [EPR20261] ab209236).

    Immunohistochemical analysis of paraffin-embedded human tonsil Hodgkin's lymphoma labeling LAG-3 with Anti-LAG-3 antibody [EPR20261] ab209236 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on immunocytes of the human Hodgkin's lymphoma [PMID: 11527700; PMID: 16757686].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    All lanes: Western blot - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (ab282638)

    Predicted band size: 57 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (ab282638), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAG-3 antibody [EPR20261] - Low endotoxin, Azide free (ab282638)

    This IHC data was generated using the same anti-LAG-3 antibody clone, EPR20261, in a different buffer formulation (Anti-LAG-3 antibody [EPR20261] ab209236).

    Immunohistochemical analysis of paraffin-embedded human tonsil Hodgkin's lymphoma labeling LAG-3 with Anti-LAG-3 antibody [EPR20261] ab209236 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on immunocytes of the human Hodgkin's lymphoma [PMID: 11527700; PMID: 16757686].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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