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AB270908

Alexa Fluor® 647 Anti-LAG-3 antibody [CAL77]

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Rabbit Recombinant Monoclonal LAG-3 antibody - conjugated to Alexa Fluor® 647. Suitable for Flow Cyt (Intra), IHC-P and reacts with Human, Mouse samples.

View Alternative Names

CD223, FDC, LAG3, Lymphocyte activation gene 3 protein, LAG-3

2 Images
Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-LAG-3 antibody [CAL77] (AB270908)
  • Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-LAG-3 antibody [CAL77] (AB270908)

Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) (top), or PBMCs treated with phytohaemagglutinin (PHA) (5 μg/ml, 48h) (bottom) with ab270908 (right) or Rabbit IgG (monoclonal) Alexa Fluor® 647 isotype control (left). PBMCs were fixed and permeabilised with BD Cytofix/Cytoperm™ for 20 min. Then incubated in 1x PBS containing 10 μg/ml anti CD16/CD32 and 10% normal goat serum followed by the antibody (ab270908) or Rabbit IgG (monoclonal) Alexa Fluor® 488 isotype (1x 106 in 100 μl at 0.04 μg/ml (1/12500 dilution)) for 30 min on ice. The cells were also stained for CD3.

Acquisition of >30000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter. Events were gated on live lymphocytes.

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-LAG-3 antibody [CAL77] (AB270908)
  • Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-LAG-3 antibody [CAL77] (AB270908)

Flow cytometry staining of C57 BL/6 mouse splenocytes (top) or C57 BL/6 mouse splenocytes treated with Concanavalin A, 3 Days, 2.5μg/mL (bottom) with ab270908 (right) or Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Cells were fixed and permeabilised with BD Cytofix/Cytoperm™ for 20 min. Cells were incubated for 30min at 22°C in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab270908 or Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 106 in 100 μl at 0.2 μg/ml (1/2500) for 30min at 22°C. The cells were simultaneously stained with CD3. Acquisition of >30000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter. Events were gated on viable lymphocytes.

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

CAL77

Isotype

IgG

Conjugation

Alexa Fluor® 647

Excitation/Emission

Ex: 650nm, Em: 665nm

Carrier free

No

Reacts with

Human, Mouse, Human, Mouse

Applications

IHC-P, Flow Cyt (Intra)

applications

Immunogen

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

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "1/2500", "FlowCytIntra-species-notes": "<p></p>", "IHCP-species-checked": "guaranteed", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>" }, "Mouse": { "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "0.2 µg/mL", "FlowCytIntra-species-notes": "<p></p>", "IHCP-species-checked": "predicted", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" } } }
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Product details

This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

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.

Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.4 Preservative: 0.02% Sodium azide Constituents: PBS, 30% Glycerol (glycerin, glycerine), 1% BSA
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|Store in the dark
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Supplementary information

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

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.

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.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

Lymphocyte activation gene 3 protein. Inhibitory receptor on antigen activated T-cells (PubMed : 20421648, PubMed : 35761082, PubMed : 7805750, PubMed : 8647185). Delivers inhibitory signals upon binding to ligands, such as MHC class II, its main ligand present at the surface of antigen-presenting cells (APCs), and FGL1, which is secreted by hepatocytes and certain types of tumor cells (PubMed : 30580966, PubMed : 32920841, PubMed : 35761082, PubMed : 39671469, PubMed : 7589152, PubMed : 8647185, PubMed : 9159144). Ligand-binding initiates a signaling that inhibits the T-cell receptor (TCR) in the immunological synapse, preventing T-cell activation (PubMed : 40101708). Mechanistically, ligand-binding promotes (1) ubiquitination of the KIEELE motif, unleashing the RRFSALE motif from the membrane and (2) leading to the formation of condensates with the TCR component CD3E, thereby disrupting the association between CD3E and LCK and preventing TCR activation (PubMed : 40101708, PubMed : 40592325). May inhibit antigen-specific T-cell activation in synergy with 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).. The LAG3-mediated inhibitory pathway is exploited by tumors to attenuate anti-tumor immunity and escape destruction by the immune system, thereby facilitating tumor survival (PubMed : 35981087, PubMed : 40101708). The blockage of the LAG3- and PDCD1-mediated pathways results in the reversal of the exhausted T-cell phenotype and the normalization of the anti-tumor response, providing a rationale for cancer immunotherapy (PubMed : 35981087, PubMed : 40101708).. 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.
See full target information LAG3
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Product promise

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