Overview

  • Product name
    Human TIM-3 ELISA Kit
    See all TIM 3 kits
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Human serum 9 2.3%
    Inter-assay
    Sample n Mean SD CV%
    Human serum 3 4.6%
  • Sample type
    Cell culture supernatant, Urine, Serum, Cell culture extracts, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    14.6 pg/ml
  • Range
    78.13 pg/ml - 5000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Urine 96 95% - 98%
    Serum 95 92% - 97%
    Cell culture extracts 104 103% - 105%
    Cell culture media 99 98% - 100%
    Heparin Plasma 88 87% - 89%
    EDTA Plasma 97 96% - 99%
    Citrate Plasma 93 93% - 94%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse, Rat, Cow
  • Product overview

    TIM-3 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of TIM-3 protein in human serum, plasma, urine, cell culture supernatant, and cell extract samples.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    T-Cell Membrane Protein 3 (TIM-3), also known as Hepatitis A Virus Cellular Receptor 2, is a type 1 transmembrane protein encoded by the HAVCR2 gene. TIM-3 contains a 181- amino acid extracellular portion that includes a Ig-like V-type domain, a 21- amino acid- transmembrane section, and a 78- amino acid cytoplasmic section. TIM-3 is a cell surface receptor expressed in Th1 lymphocytes, regulatory T-cells, innate immune cells including dendritic cells, NK cells, and monocytes. Binding to Galectin-9 induces intracellular calcium influx which triggers apoptosis. Other ligands are phosphatidyl serine, High Mobility Group Protein 1 (HMGB1), and Carcinoembryonic Antigen Related Cell Adhesion Molecule 1 (CEACAM1). TIM-3 can be cleaved to a soluble form from the cell surface by ADAM10 or ADAM17 mediated lysis. The soluble form of TIM-3 has been found to be elevated in the plasma of patients with HIV/AIDS, Hepatitis, and graft-versus-host disease.


    Sensitivity:


    Samples diluted in Sample Diluent NS: 14.6 pg/ml


    Samples diluted in 1X Cell Extraction Buffer PTR: 23.6 pg/ml

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human TIM-3 Capture Antibody 1 x 600µl
    10X Human TIM-3 Detector Antibody 1 x 600µl
    Human TIM-3 Lyophilized Recombinant Protein 2 vials
    Antibody Diluent 5BR 1 x 6ml
    10X Wash Buffer PT (ab206977) 1 x 20ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    TMB Development Solution 1 x 12ml
    Stop Solution 1 x 12ml
    Sample Diluent NS 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Plate Seals 1 unit
  • Research areas
  • 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
  • Alternative names
    • CD366
    • FLJ14428
    • HAVcr-2
    • Havcr2
    • HAVR2_HUMAN
    • Hepatitis A virus cellular receptor 2
    • Kidney injury molecule 3
    • KIM 3
    • KIM3
    • T cell immunoglobulin and mucin domain containing 3
    • T cell immunoglobulin mucin 3
    • T-cell immunoglobulin and mucin domain-containing protein 3
    • T-cell immunoglobulin mucin family member 3
    • T-cell immunoglobulin mucin receptor 3
    • T-cell membrane protein 3
    • Tim 3
    • TIM-3
    • TIM3
    • TIMD-3
    • TIMD3
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab231932 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • ELISA Protocol Summary
  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of TIM-3 were measured in duplicates, interpolated from the TIM-3 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (heparin) 12.5%, plasma (citrate) 12.5%, plasma (EDTA) 50%, RPMI media 25%, and urine 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

  • The concentrations of TIM-3 were measured in duplicate and interpolated from the TIM-3 standard curve and corrected for sample dilution. Undiluted samples are 100 µg/mL. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

  • Human PBMCs were cultured and either stimulated with 1.5% PHA-M or mock stimulated for 2 days. The concentrations of TIM-3 were measured in duplicate and interpolated from the TIM-3 standard curve and corrected for sample dilution. Undiluted samples are 100 µg/mL. The average interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TIM-3 concentration was determined to be 412 pg/mL in stimulated PBMCs and 92 pg/mL in unstimulated PBMCs.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TIM-3 concentration was determined to be 202 pg/mL with a range of 122 – 321 pg/mL.

Protocols

References

ab231932 has not yet been referenced specifically in any publications.

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