Natural and recombinant mouse TIM-1. The antibody components of this kit were raised against the short splice form of mouse TIM-1 (isoform 2) which has a 23 amino acid deletion following Pro182. This immunoassay detects both the short and long forms of mouse TIM-1.
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
No significant interference observed with available related molecules.
The Quantikine Mouse TIM-1 Immunoassay is a 3.5 hour solid phase ELISA designed to measure mouse TIM-1 in cell culture supernates, serum, plasma, and urine. It contains NS0-expressed recombinant mouse TIM-1 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant factor accurately. Results obtained using natural mouse TIM-1 showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse TIM-1.
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
The recovery of TIM-1 spiked into various matrices was evaluated.
Average % Recovery
Cell Culture Media (n=4)
EDTA Plasma (n=4)
Heparin Plasma (n=4)
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of mouse TIM-1 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
TIM-1 (T cell immunoglobulin and mucin domain 1), also known as KIM-1 and HAVcr1, is expressed on many immune cell types and epithelial cells. It binds to phosphatidylserine (PS), LMIR5/CD300b, TIM-1 (homophilic), TIM-4, IgA, and the glycoproteins of a number of enveloped viruses. Its interaction with PS enables TIM-1 to mediate the phagocytosis and clearance of apoptotic cells. TIM-1 signaling costimulates T cell activation and enhances Th2 cytokine production. TIM-1 serves as a cellular entry receptor for hepatitis A virus, Ebolavirus and Marburgvirus. Polymorphisms are associated with susceptibility to atopy, autoimmunity, and severe hepatitis A virus infection in humans. A soluble form of TIM-1 is elevated in the urine during nephropathy.
Refer to the product for complete assay procedure.
The conjugate must remain at 2-8 °C during use. Bring all other reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
50 µL Assay Diluent
Add 50 µL of Assay Diluent to each well.
50 µL Standard, Control, or Sample
Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
100 µL cold Conjugate
Add 100 µL of cold Conjugate to each well. Cover with a new plate sealer, and incubate at 2-8 °C for 1 hour on the benchtop.
Aspirate and wash 5 times.
100 µL Substrate Solution
Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
100 µL Stop Solution
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.
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