Description
Description: This assay employs a two-site sandwich ELISA to quantitate KLHL4 in samples. An antibody specific for KLHL4 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any KLHL4 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for KLHL4 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 KLHL4 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Overview: The KLHL4 transcript represents an approximately 3.6-kb mRNA and encodes a 718-amino acid protein. PCR analysis demonstrated alternative use of exons 11a and 11d, with the exon 11d transcript encoding a 719-amino acid protein. KLHL4 contains 6 tandem repeats (Kelch repeats) at the C terminus and a POZ/BTB protein-binding domain toward the N terminus, characteristic of Kelch family members. The POZ/BTB domain is a 120-amino acid motif generally located in the N terminus of actin-binding proteins and believed to be important in protein-protein interactions, allowing homomeric or heteromeric dimerization. Kelch-like proteins usually have distinct N-terminal ends, but KLHL4 shows 88% similarity over the first 25 amino acids with KLHL1