Overview

  • Product name

    Anti-TIM 3 antibody [EPR22241]
    See all TIM 3 primary antibodies
  • Description

    Rabbit monoclonal [EPR22241] to TIM 3
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment within Human TIM 3 aa 200-300. The exact sequence is proprietary.
    Database link: Q8TDQ0

  • Positive control

    • WB: Daudi and RAW 264.7 whole cell lysates (+/- treatment with PNGase F). IP: Daudi whole cell lysate. IHC-P: Human liver and lung cancer tissue; mouse spleen tissue; rat spleen tissue.
  • 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

    EPR22241
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab241332 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 50-60 kDa (predicted molecular weight: 33 kDa).
IP 1/30.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt or ICC/IF.
  • 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

    • Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling TIM 3 with ab241332 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic and membranous staining on both infiltrated immunocytes and tumor cells of human lung cancer (PMID: 22586058) 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).

    • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling TIM 3 with ab241332 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic and membranous staining on subsets of immune cells of mouse spleen 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).

    • Immunohistochemical analysis of paraffin-embedded human liver tissue labeling TIM 3 with ab241332 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic and membranous staining on Kupffer cells of human liver (PMID: 27192565) 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).

    • Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling TIM 3 with ab241332 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic and membranous staining on subsets of immune cells of rat spleen 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).

    • All lanes : Anti-TIM 3 antibody [EPR22241] (ab241332) at 1/1000 dilution

      Lane 1 : Daudi (human Burkitt's lymphoma cell line) whole cell lysate at 20 µg
      Lane 2 : Daudi whole cell lysate (treated with PNGase F) at 20 mg/ml
      Lane 3 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 20 µg
      Lane 4 : RAW 264.7 whole cell lysate (treated with PNGase F) at 20 µg

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Developed using the ECL technique.

      Predicted band size: 33 kDa
      Observed band size: 40 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 50-60 kDa (possible glycosylated form)


      Exposure time: 70 seconds


      Blocking and dilution buffer: 5% NFDM/TBST.

      Lanes 3 & 4 were developed using a higher sensitivity ECL substrate.

    • TIM 3 was immunoprecipitated from 0.35 mg of Daudi (human Burkitt's lymphoma cell line) whole cell lysate with ab241332 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab241332 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

      Lane 1: Daudi lysate 10 μg (Input).
      Lane 2: ab241332 IP in Daudi whole cell lysate (+).
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab241332 in Daudi whole cell lysate (-).

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 3 minutes.

    References

    ab241332 has not yet been referenced specifically in any publications.

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