Human TIM-1/KIM-1 ELISA Kit

Principle of the Assay

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for TIM-1/KIM-1/HAVCR has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any TIM-1/KIM-1/HAVCR present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for TIM-1/KIM-1/HAVCR is added to the wells and binds to the combination of capture antibody-TIM-1/KIM-1/HAVCR in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps a substrate is added. A coloured product is formed in proportion to the amount of TIM-1/KIM-1/HAVCR present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven TIM-1/KIM-1/HAVCR standard dilutions and TIM-1/KIM-1/HAVCR sample concentration determined.

For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.

Target Information

T cell immunoglobulin and mucin domain 1 (TIM-1), also known as Kidney injury molecule 1 (KIM-1) and Hepatitis A virus cellular receptor 1 (HAVcr1), is a member of the TIM family which is involved in the regulation of innate and adaptive immune responses. TIM-1 is a type I transmembrane protein that contains an N-terminal immunoglobulin-like domain, a mucin domain with O- and N-linked glycosylations, a transmembrane segment, and a cytoplasmic signaling domain. Multiple TIM-1 variants can be produced due to polymorphisms or alternative splicing resulting in deletions in the mucin domain. Some of these polymorphisms are associated with susceptibility to atopy, autoimmunity, and severe hepatitis A viral infection in humans. Within the extracellular domain, human TIM-1 shares 41% amino acid sequence identity with mouse and rat TIM-1.

In vivo, TIM-1 is expressed on splenic B cells and is a marker for the identification of IL-10+ regulatory B cells. TIM-1 is also expressed on CD4+ T cells, mast cells, invariant NKT (iNKT) cells, dendritic cells, kidney epithelium and a broad range of mucosal epithelium. The expression of TIM-1 is upregulated on activated Th2 cells, after dendritic cell maturation, and on kidney tubular epithelial cells after injury. Metalloproteinase-mediated cleavage of TIM-1 at the membrane-proximal region results in the release of a soluble form of TIM-1 which is detectable in the urine and in circulation. Urinary TIM-1 is highly elevated in nephropathy and may be a useful biomarker for renal damage.

TIM-1 has been reported to be a receptor for a number of ligands, including phosphatidylserine, leukocyte mono-immunoglobulin-like receptor 5 (LMIR5/CD300b), TIM-1 (homophilic), TIM-4, IgA, and the glycoproteins of a number of enveloped viruses. Its interaction with phosphatidylserine enables TIM-1 to mediate the phagocytosis of apoptotic cells. In TIM-1-bearing iNKT cells, interaction with apoptotic cells can also result in iNKT cell activation, proliferation, and cytokine production. Interactions between cell-surface or soluble TIM-1 with LMIR5 is proposed to induce LMIR5-mediated activation of myeloid cells including macrophages/monocytes, mast cells, neutrophils, and dendritic cells. These interactions contribute to tissue homeostasis and damage during kidney injury. Ligandinduced TIM-1 signaling costimulates T cell activation and enhances Th2 cytokine production. In humans, TIM-1 serves as a cellular entry receptor for various viruses, including hepatitis A virus, Ebolavirus and Marburgvirus.

GENE ID 26762
SWISS PROT Q96D42
SYNONYMS TIM; KIM1; TIM1; CD365; HAVCR; KIM-1; TIM-1; TIMD1; TIMD-1; HAVCR-1

Materials Supplied

Kit Components 96 Wells Quantity/Size
Aluminium pouches with a Microwell Plate coated with monoclonal antibody to human TIM-1/KIM-1/HAVCR (8﹡12) 1 plate
Human TIM-1/KIM-1/HAVCR Standard lyophilized, 1000 pg/ml upon reconstitution 2 vials
Concentrated Biotin-Conjugate anti-human TIM-1/KIM-1/HAVCR monoclonal antibody 2 vials
Streptavidin-HRP solution 2 vials
Standard /sample Diluent 1 bottle
Biotin-Conjugate antibody Diluent 1 bottle
Streptavidin-HRP Diluent 1 bottle
Wash Buffer Concentrate 20x (PBS with 1% Tween-20) 1 bottle
Substrate Solution 1 vial
Stop Solution 1 vial
Adhesive Films 4 pieces
Product data sheet 1 copy

Storage

Storage Store at 2 - 8°C

Performance Characteristics

REPEATABILITY The coefficient of variation of both intra-assay and inter-assay were less than 10%.
SENSITIVITY The minimum detectable dose was 7pg/mL.
ASSAY RANGE 15.6 - 1000 pg/mL
SPECIFICITY This assay recognizes both natural and recombinant human TIM-1/KIM-1/HAVCR. The factors listed below were prepared at 50ng/ml in Standard /sample Diluent and assayed for cross-reactivity and no significant cross-reactivity or interference was observed.
Factors assayed for cross-reactivity
Recombinant human TIM-3, TIM-4
Recombinant mouse TIM-1, TIM-4
Recombinant rat TIM-1