Rat TIM-1/KIM-1/HAVCR Quantikine ELISA Kit Summary

Product Summary

The Quantikine Rat TIM-1 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure rat TIM-1 in cell culture supernates, tissue homogenates, serum, plasma, and urine. It contains NS0-expressed recombinant rat TIM-1 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural rat TIM-1 showed linear 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 mass values for naturally occurring rat TIM-1.

Preparation and Storage

Background: TIM-1/KIM-1/HAVCR

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.

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.