Description
Description: This assay employs a two-site sandwich ELISA to quantitate MARK4 in samples. An antibody specific for MARK4 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any MARK4 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for MARK4 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 MARK4 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Overview: Sequence analysis predicted that the 752-amino acid MARKL1 protein, 62% identical to MARK3, contains an N-terminal serine/threonine kinase domain, a central ubiquitin-associated domain, and a C-terminal KA1 -associated kinase domain. A smaller transcript, MARKL1S, lacking exon 16 and the KA1-associated kinase domain in the 688-amino acid protein, was also detected exclusively in brain. RT-PCR analysis detected upregulated expression in nearly all clinical hepatocellular carcinoma cells. Northern blot analysis revealed ubiquitous expression of a 3.6-kb transcript, with highest expression in testis. Immunofluorescence microscopy demonstrated homogeneous cytoplasmic expression. Colony-forming assays showed that MARKL1 antisense reduced the growth of SNU475 cells