Recombinant

Recombinant Anti-LAG-3 antibody [CAL26] (ab237719)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [CAL26] to LAG-3
  • Suitable for: WB, Flow Cyt, IP, IHC-P
  • Reacts with: Human

Overview

  • Product name

    Anti-LAG-3 antibody [CAL26]
    See all LAG-3 primary antibodies

  • Description

    Rabbit monoclonal [CAL26] to LAG-3

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, Flow Cyt, IP, IHC-Pmore details
    Unsuitable for: ICC/IF

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is considered to be commercially sensitive.

  • Positive control

    • WB: LAG-3-GFP transfected HEK-293 whole cell lysate; human PBMC whole cell lysate. IHC-P: Human tonsil tissue. Flow cyt: Human PBMCs treated with 10µg/ml PHA for 48 hours. IP: LAG-3-GFP transfected HEK-203& whole cell lysate.

  • General notes

    This product was previously labelled as Lymphocyte Activation Gene 3.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab237719 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Detects a band of approximately 16, 70 kDa (predicted molecular weight: 57 kDa).
Flow Cyt 1/250.
IP 1/20.
IHC-P Use a concentration of 0.3 µg/ml. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for ICC/IF.

    • Target

    • Function

      Involved in lymphocyte activation. Binds to HLA class-II antigens.

    • Tissue specificity

      On cell surface of activated NK and T-lymphocytes.

    • Sequence similarities

      Contains 3 Ig-like C2-type (immunoglobulin-like) domains.
      Contains 1 Ig-like V-type (immunoglobulin-like) domain.

    • Cellular localization

      Membrane.
    • Information by UniProt

    • Database links

      • Alternative names

        • CD223 antibody
        • CD223 antigen antibody
        • FDC protein antibody
        • LAG-3 antibody
        • Lag3 antibody
        • LAG3_HUMAN antibody
        • Lymphocyte activating 3 antibody
        • Lymphocyte activation gene 3 antibody
        • Lymphocyte activation gene 3 protein antibody
        • Protein FDC antibody
        see all

      Images

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAG-3 antibody [CAL26] (ab237719)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAG-3 antibody [CAL26] (ab237719)

        Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling LAG-3 with ab237719 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on the human tonsil is observed. 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.

        Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

      • Immunoprecipitation - Anti-LAG-3 antibody [CAL26] (ab237719)
        Immunoprecipitation - Anti-LAG-3 antibody [CAL26] (ab237719)

        LAG-3 was immunoprecipitated from 0.35 mg of LAG-3-GFP transfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen)  whole cell lysate with ab237719 at 1/20 dilution. Western blot was performed from the immunoprecipitate using ab237719 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

        Lane 1: LAG-3-GFP transfected HEK-293T whole cell lysate 10 μg (Input).
        Lane 2: ab237719 IP in LAG-3-GFP transfected HEK-293T whole cell lysate.
        Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab237719 in LAG-3-GFP transfected HEK-293T whole cell lysate.

        Blocking and dilution buffer and concentration: 5% NFDM/TBST.
        Exposure time: 30 seconds.

      • Flow Cytometry - Anti-LAG-3 antibody [CAL26] (ab237719)
        Flow Cytometry - Anti-LAG-3 antibody [CAL26] (ab237719)

        Flow cytometric analysis of 2% paraformaldehyde-fixed, 0.1% Tween-20 permeabilized human peripheral blood mononuclear cells ((PBMC) (treated with 10μg/ml PHA for 48 hours) cell line labeling LAG-3 with ab237719 at 1/250 dilution (Right) compared with Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (Left).

        Goat Anti-Rabbit IgG Fc (Alexa Fluor® 488) preadsorbed (ab150097), at 1/5000 dilution was used as the secondary antibody.

        Cells were surface stained with anti-CD3 conjugated to Alexa Fluor® 647. Then fixed with 2% PFA for 10min followed by intracellular staining rabbit IgG (Left) or ab237719 (Right).

      • Western blot - Anti-LAG-3 antibody [CAL26] (ab237719)
        Western blot - Anti-LAG-3 antibody [CAL26] (ab237719)
        All lanes : Anti-LAG-3 antibody [CAL26] (ab237719) at 1/1000 dilution

        Lane 1 : Untransfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
        Lane 2 : LAG-3-GFP overexpression HEK-293T whole cell lysate

        Lysates/proteins at 20 µg per lane.

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

        Predicted band size: 57 kDa
        Observed band size: 16,70 kDa why is the actual band size different from the predicted?


        Exposure time: 3 minutes


        Blocking and dilution buffer: 5% NFDM/TBST.

        This antibody also detects a 16kDa cleaved fragment. (PMID: 15557174).

      • Western blot - Anti-LAG-3 antibody [CAL26] (ab237719)
        Western blot - Anti-LAG-3 antibody [CAL26] (ab237719)
        All lanes : Anti-LAG-3 antibody [CAL26] (ab237719) at 1/1000 dilution

        Lane 1 : Human peripheral blood mononuclear cell (PBMC), whole cell lysate
        Lane 2 : Human peripheral blood mononuclear cell (PBMC) treated with 10µg/ml PHA for 48 hours, whole cell lysate
        Lane 3 : Human lymphoma lysate

        Lysates/proteins at 20 µg per lane.

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

        Predicted band size: 57 kDa
        Observed band size: 16,70 kDa why is the actual band size different from the predicted?


        Exposure time: 3 minutes


        Blocking and dilution buffer: 5% NFDM/TBST.

        This antibody also detects a 16kDa cleaved fragment. (PMID: 15557174).

         

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAG-3 antibody [CAL26] (ab237719)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAG-3 antibody [CAL26] (ab237719)

        Formalin-fixed, paraffin-embedded human tonsil tissue stained for LAG-3 using ab237719 at 0.3 μg/ml in immunohistochemical analysis.

      • Anti-LAG-3 antibody [CAL26] (ab237719)
        Anti-LAG-3 antibody [CAL26] (ab237719)

      Protocols

      To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

      Click here to view the general protocols

      References (0)

      Publishing research using ab237719? Please let us know so that we can cite the reference in this datasheet.

      ab237719 has not yet been referenced specifically in any publications.

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