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Rabbit Recombinant Monoclonal LAG-3 antibody. Suitable for IP, Flow Cyt, WB, ICC/IF, IHC-P and reacts with Human samples. Cited in 13 publications.


Images

Flow Cytometry - Anti-LAG-3 antibody [EPR20261] (AB209236), expandable thumbnail
  • Western blot - Anti-LAG-3 antibody [EPR20261] (AB209236), expandable thumbnail
  • Flow Cytometry - Anti-LAG-3 antibody [EPR20261] (AB209236), expandable thumbnail
  • Western blot - Anti-LAG-3 antibody [EPR20261] (AB209236), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAG-3 antibody [EPR20261] (AB209236), 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
IPFlow CytWBICC/IFIHC-P
Human
Tested
Tested
Tested
Tested
Tested

Tested
Tested

Species

Human

Dilution info

1/30

Notes

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

Tested
Tested

Species

Human

Dilution info

1/500

Notes

-

Tested
Tested

Species

Human

Dilution info

1/500

Notes

-

Tested
Tested

Species

Human

Dilution info

1/100

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000

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.

Associated Products

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Target data

Function

Lymphocyte activation gene 3 protein: Inhibitory receptor on antigen activated T-cells (PubMed:7805750, PubMed:8647185, PubMed:20421648). 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:7805750, PubMed:8647185, PubMed:20421648). 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 proteinMay 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

Lymphocyte activation gene 3 protein, LAG-3, LAG3, FDC

Recommended products

Rabbit Recombinant Monoclonal LAG-3 antibody. Suitable for IP, Flow Cyt, WB, ICC/IF, IHC-P and reacts with Human samples. Cited in 13 publications.

Alternative names

Lymphocyte activation gene 3 protein, LAG-3, LAG3, FDC

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

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

EPR20261

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

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.

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.

11 product images

  • Flow Cytometry - Anti-LAG-3 antibody [EPR20261] (ab209236), expandable thumbnail

    Flow Cytometry - 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 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 LAG3. Gated on total viable cells.

  • Western blot - Anti-LAG-3 antibody [EPR20261] (ab209236), expandable thumbnail

    Western blot - Anti-LAG-3 antibody [EPR20261] (ab209236)

    Blocking/Diluting buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-LAG-3 antibody [EPR20261] (ab209236) at 1/5000 dilution

    Lane 1: 293T (Human embryonic kidney epithelial cell) transfected with an empty vector (vector control), containing a GFP tag, whole cell lysate at 15 µg

    Lane 2: 293T transfected with LAG3 expression vector containing a GFP tag, whole cell lysate at 15 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

    Predicted band size: 57 kDa

    Exposure time: 5s

  • Flow Cytometry - Anti-LAG-3 antibody [EPR20261] (ab209236), expandable thumbnail

    Flow Cytometry - 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 LAG-3 construct labeling LAG-3 with 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).

  • Western blot - Anti-LAG-3 antibody [EPR20261] (ab209236), expandable thumbnail

    Western blot - 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 – ab209236 observed at 54-70 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
    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] (ab209236) 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

    This IHC data was generated using the same anti-LAG-3 antibody clone, EPR20261, in a different buffer formulation (cat# 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 – ab209236 observed at 54-70 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAG-3 antibody [EPR20261] (ab209236), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAG-3 antibody [EPR20261] (ab209236)

    Immunohistochemical analysis of paraffin-embedded human tonsil Hodgkin's lymphoma labeling LAG-3 with 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-LAG-3 antibody [EPR20261] (ab209236), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - 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 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 LAG3 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] (ab209236), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAG-3 antibody [EPR20261] (ab209236)

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling LAG-3 with 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.

  • Immunoprecipitation - Anti-LAG-3 antibody [EPR20261] (ab209236), expandable thumbnail

    Immunoprecipitation - Anti-LAG-3 antibody [EPR20261] (ab209236)

    LAG-3 was immunoprecipitated from 0.35 mg of HEK-293T (Human epithelial cell line from embryonic kidney) transfected with a GFP-tagged human LAG3 construct whole cell lysate with ab209236 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab209236 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HEK-293T transfected with a GFP-tagged human LAG3 construct whole cell lysate 10 μg (Input).

    Lane 2: ab209236 IP in HEK-293T transfected with a GFP-tagged human LAG3 construct whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab209236 in HEK-293T transfected with a GFP-tagged human LAG3 construct whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 1 second.

    All lanes: Immunoprecipitation - Anti-LAG-3 antibody [EPR20261] (ab209236)

    Predicted band size: 57 kDa

  • Western blot - Anti-LAG-3 antibody [EPR20261] (ab209236), expandable thumbnail

    Western blot - Anti-LAG-3 antibody [EPR20261] (ab209236)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-LAG-3 antibody [EPR20261] (ab209236) at 1/1000 dilution

    All lanes: Human LAG3 Fc chimera recombinant protein (aa23-450) at 0.01 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

    Predicted band size: 57 kDa

    Observed band size: 90 kDa

    Exposure time: 1s

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAG-3 antibody [EPR20261] (ab209236), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAG-3 antibody [EPR20261] (ab209236)

    Tissue Microarrays stained for "Anti-LAG-3 antibody [EPR20261]" using "ab209236"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). The sections were incubated with ab209236 at +4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

  • Flow Cytometry - Anti-LAG-3 antibody [EPR20261] (ab209236), expandable thumbnail

    Flow Cytometry - Anti-LAG-3 antibody [EPR20261] (ab209236)

    Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) (top) or PBMCs treated with 5 μg/ml phytohaemagglutinin (PHA) for 72 hours (bottom), with ab209236 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). PBMCs were incubated for 30 min on ice in 1x PBS containing 10 µg/ml human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab209236 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 106 in 100 µl at 5.0 μg/ml (1/402)) for 30min on ice. The cells were simultaneously stained with CD3.

    The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice

    Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable cells.

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