Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate TIMD4 in samples. An antibody specific for TIMD4 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyTIMD4 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for TIMD4 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of TIMD4 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:TIM4 has an N-terminal IgV domain, followed by a long mucin stalk and a C-terminal cytoplasmic tail. The IgV domain contains an N-linked glycosylation site and a potential integrin-binding RGD motif, and the mucin stalk contains 38 potential O-linked glycosylation sites and a second N-linked glycosylation site near the membrane. The cytoplasmic domain of TIM4 lacks a conserved tyrosine kinase phosphorylation site found in other TIM family members.
The antigen recognized by the antibody was identified by expression cloning as a type I transmembrane protein called Tim4. Tim4 was expressed in Mac1+ cells in various mouse tissues, including spleen, lymph nodes, and fetal liver. Tim4 bound apoptotic cells by recognizing phosphatidylserine via its immunoglobulin domain.
