Description
Principle of the assay: mouse KIM1 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for KIM1 has been precoated onto 96-well plates. Standards(NSO, Y22-T212) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for KIM1 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the mouse KIM1 amount of sample captured in plate.
Background: KIM1(TIM-1), also known as Hepatitis A virus cellular receptor 1, is a protein that in Mouses is encoded by the HAVCR1 gene. Infection of canine osteogenic sarcoma cells expressing HAVCR1 with HAV led Feigelstock et al. (1998) to conclude that the protein is indeed a receptor for the virus. Immunofluorescence microscopy demonstrated internalization of HAV by dog cells expressing HAVCR1. Using a monoclonal antibody to mouse Tim1, Umetsu et al. (2005) showed that Tim1 was expressed after activation of naive T cells and on T cells differentiated in Th2-polarizing conditions. By homology of synteny with the mouse Tim1 gene and database analysis, McIntire et al. (2001) mapped the HAVCR1 gene to 5q33.2. The standard used in this kit is recombinant mouse KIM-1, constituting from Y22-T212 amino acids, and 191amino acids with the molecular mass of 21.8KDa