Overview

  • Product name

    Anti-TIM 3 antibody [EPR20767]
    See all TIM 3 primary antibodies
  • Description

    Rabbit monoclonal [EPR20767] to TIM 3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, Flow Cyt, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human TIM 3 aa 1-200. The exact sequence is proprietary.
    Database link: Q8TDQ0

  • Positive control

    • WB: Fc-tagged human TIM 3 recombinant protein (aa22-200); Daudi whole cell lysate. Flow Cyt: Human peripheral blood mononuclear cells (PBMCs). IP: Daudi whole cell lysate.
  • General 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 rabbit monoclonal antibody.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20767
  • Isotype

    IgG
  • Light chain type

    kappa
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab210543 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
IP 1/30.
Flow Cyt 1/500.
WB 1/1000. Detects a band of approximately 50 kDa (predicted molecular weight: 33 kDa).

Target

  • Function

    Cell surface receptor implicated in modulating innate and adaptive immune responses. Generally accepted to have an inhibiting function. Reports on stimulating functions suggest that the activity may be influenced by the cellular context and/or the respective ligand (PubMed:24825777). Regulates macrophage activation (PubMed:11823861). Inhibits T-helper type 1 lymphocyte (Th1)-mediated auto- and alloimmune responses and promotes immunological tolerance (PubMed:14556005). In CD8+ cells attenuates TCR-induced signaling, specifically by blocking NF-kappaB and NFAT promoter activities resulting in the loss of IL-2 secretion. The function may implicate its association with LCK proposed to impair phosphorylation of TCR subunits, and/or LGALS9-dependent recruitment of PTPRC to the immunological synapse (PubMed:24337741, PubMed:26492563). In contrast, shown to activate TCR-induced signaling in T-cells probably implicating ZAP70, LCP2, LCK and FYN (By similarity). Expressed on Treg cells can inhibit Th17 cell responses (PubMed:24838857). Receptor for LGALS9 (PubMed:16286920, PubMed:24337741). Binding to LGALS9 is believed to result in suppression of T-cell responses; the resulting apoptosis of antigen-specific cells may implicate HAVCR2 phosphorylation and disruption of its association with BAG6. Binding to LGALS9 is proposed to be involved in innate immune response to intracellular pathogens. Expressed on Th1 cells interacts with LGALS9 expressed on Mycobacterium tuberculosis-infected macrophages to stimulate antibactericidal activity including IL-1 beta secretion and to restrict intracellular bacterial growth (By similarity). However, the function as receptor for LGALS9 has been challenged (PubMed:23555261). Also reported to enhance CD8+ T-cell responses to an acute infection such as by Listeria monocytogenes (By similarity). Receptor for phosphatidylserine (PtSer); PtSer-binding is calcium-dependent. May recognize PtSer on apoptotic cells leading to their phagocytosis. Mediates the engulfment of apoptotic cells by dendritic cells. Expressed on T-cells, promotes conjugation but not engulfment of apoptotic cells. Expressed on dendritic cells (DCs) positively regulates innate immune response and in synergy with Toll-like receptors promotes secretion of TNF-alpha. In tumor-imfiltrating DCs suppresses nucleic acid-mediated innate immune repsonse by interaction with HMGB1 and interfering with nucleic acid-sensing and trafficking of nucleid acids to endosomes (By similarity). Expressed on natural killer (NK) cells acts as a coreceptor to enhance IFN-gamma production in response to LGALS9 (PubMed:22323453). In contrast, shown to suppress NK cell-mediated cytotoxicity (PubMed:22383801). Negatively regulates NK cell function in LPS-induced endotoxic shock.
  • Tissue specificity

    Expressed in T-helper type 1 (Th1) lymphocytes. Expressed on regulatory T (Treg) cells after TCR stimulation. Expressed in dendritic cells and natural killer (NK) cells. Expressed in epithelial tissues. Expression is increased on CD4+ and CD8+ T-cells in chronic hepatitis C virus (HCV) infection. In progressive HIV-1 infection, expression is up-regulated on HIV-1-specific CD8 T-cells.
  • Involvement in disease

    May be involved in T-cell exhaustion associated with chronic viral infections such as with human immunodeficiency virus (HIV) and hepatitic C virus (HCV).
  • Sequence similarities

    Belongs to the immunoglobulin superfamily. TIM family.
    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • Post-translational
    modifications

    O-glycosylated with core 1 or possibly core 8 glycans.
    Phosphorylated on tyrosine residues; modestly increased after TCR/CD28 stimulation. Can be phosphorylated in the cytoplasmatic domain by FYN (By similarity). Phosphorylation at Tyr-265 is increased by stimulation with ligand LGALS9.
  • Cellular localization

    Membrane. Cell junction. Localizes to the immunological synapse between CD8+ T-cells and target cells.
  • Information by UniProt
  • Database links

  • Alternative names

    • CD366 antibody
    • FLJ14428 antibody
    • HAVcr-2 antibody
    • Havcr2 antibody
    • HAVR2_HUMAN antibody
    • Hepatitis A virus cellular receptor 2 antibody
    • Kidney injury molecule 3 antibody
    • KIM 3 antibody
    • KIM3 antibody
    • T cell immunoglobulin and mucin domain containing 3 antibody
    • T cell immunoglobulin mucin 3 antibody
    • T-cell immunoglobulin and mucin domain-containing protein 3 antibody
    • T-cell immunoglobulin mucin family member 3 antibody
    • T-cell immunoglobulin mucin receptor 3 antibody
    • T-cell membrane protein 3 antibody
    • Tim 3 antibody
    • TIM-3 antibody
    • TIM3 antibody
    • TIMD-3 antibody
    • TIMD3 antibody
    see all

Images

  • Flow cytometric analysis of human peripheral blood mononuclear cells (PBMCs) labeling TIM 3 with ab210543 at 1/500 dilution (right panel) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

    Co-staining with anti-CD14 (y-axis). CD14+ PBMCs were positive for TIM 3 (PMID 22426209). Gated on total viable cells.

  • Anti-TIM 3 antibody [EPR20767] (ab210543) at 1/1000 dilution + Fc-tagged human TIM 3 recombinant protein (aa22-200) at 0.01 µg

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

    Developed using the ECL technique.

    Predicted band size: 33 kDa
    Observed band size: 75 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-TIM 3 antibody [EPR20767] (ab210543) at 1/1000 dilution + Daudi (human Burkitt's lymphoma cell line) whole cell lysate at 10 µg

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

    Developed using the ECL technique.

    Predicted band size: 33 kDa
    Observed band size: 50 kDa why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Strong signal detected in Daudi cells after 15s exposure. Please note that under our experimental conditions (overexposure >3min), a faint band at ~70kDa was detected in Jurkat T cells (previously reported as a TIM 3 negative cell line PMID: 21807895).

  • TIM 3 was immunoprecipitated from 0.35 mg of Daudi (human Burkitt's lymphoma cell line) whole cell lysate with ab210543 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab210543 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Lane 1: Daudi whole cell lysate 10 µg (Input).
    Lane 2: ab210543 IP in Daudi whole cell lysate. 
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab210543 in Daudi whole cell lysate.

    Exposure time: 10 seconds.

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

References

ab210543 has not yet been referenced specifically in any publications.

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