Human Serum TIM-1/KIM-1/HAVCR Quantikine ELISA Kit Summary Assay Type | Solid Phase Sandwich ELISA | Format | 96-well strip plate | Assay Length | 4.5 hours | Sample Type & Volume Required Per Well | Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL) | Sensitivity | 3.63 pg/mL | Assay Range | 10.9 - 700 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma) | Specificity | Natural and recombinant human TIM-1 | Cross-reactivity | < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested | Interference | No significant interference observed with available related molecules. |
Product Summary The Quantikine® Human Serum TIM-1/KIM-1/HAVCR Immunoassay is a 4.5 hour solid phase ELISA designed to measure human TIM-1 in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human TIM-1 and antibodies raised against the recombinant factor. Natural human TIM-1 showed dose-response curves that were parallel to the standard curves obtained using the Quantikine® kit standards. These results indicate that this kit can be used to determine relative levels of natural human TIM-1. Preparation and Storage Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. | Storage | Store the unopened product at 2 - 8 °C. Do not use past expiration date. |
Background: TIM-1/KIM-1/HAVCRT 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 (1, 2). 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 (3, 4). Multiple TIM-1 variants can be produced due to polymorphisms or alternative splicing resulting in deletions in the mucin domain (3). Some of these polymorphisms are associated with susceptibility to atopy, autoimmunity, and severe hepatitis A virus infection in humans (5). 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 (6, 7). 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 (4, 8-15). The expression of TIM-1 is upregulated on activated Th2 cells, after dendritic cell maturation, and on kidney tubular epithelial cells after injury (4, 9, 13, 14, 16, 17). 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 (18, 19). Urinary TIM-1 is highly elevated in nephropathy and may be a useful biomarker for renal damage (16, 20 - 25). 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 (5, 15, 26-33). Its interaction with phosphatidylserine enables TIM-1 to mediate the phagocytosis of apoptotic cells (26-28). In TIM-1-bearing iNKT cells, interaction with apoptotic cells can also result in iNKT cell activation, proliferation, and cytokine production (11). 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 (29). These interactions contribute to tissue homeostasis and damage during kidney injury (29). Ligandinduced TIM-1 signaling costimulates T cell activation and enhances Th2 cytokine production (9, 31, 34). In humans, TIM-1 serves as a cellular entry receptor for various viruses, including hepatitis A virus, Ebolavirus and Marburgvirus (15, 33). Long Name: | T Cell Immunoglobulin Mucin-1 | Entrez Gene IDs: | 26762 (Human); 171283 (Mouse); 286934 (Rat); 102141332 (Cynomolgus Monkey) | Alternate Names: | CD365; HAVCR1; HAVCR-1; HAVCRT cell immunoglobin domain and mucin domain protein 1; hepatitis A virus cellular receptor 1; Kidney injury molecule 1; KIM1; KIM-1; T-cell immunoglobulin and mucin domain-containing protein 1; TIM1; TIM-1; TIM-1TIM; TIM1TIMD-1; TIMD1T-cell membrane protein 1 |
Assay Procedure Refer to the product datasheet for the complete assay procedure. Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate. 1. Prepare all reagents, standard dilutions, and samples as directed in the product insert. 2. Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal. 3. Add 50 μL of Assay Diluent to each well. 4. Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours. 5. Aspirate each well and wash, repeating the process 4 times for a total of 5 washes. 6. Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours. 7. Aspirate and wash 5 times. 8. Add 100 μL Substrate Solution to each well. 9. Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm. |
