Human Hyaluronan ELISA Kit
Hyaluronan, also known as hyaluronic acid (HA) or sodium hyaluronate, is a naturally occurring linear polymer. Hyaluronan is a glycosaminoglycan (GAG) that is ubiquitously present in the extracellular matrix of all vertebrates and is also present in the capsule of some strains of Streptococci. Mammalian hyaluronan is synthesized by one of three distinct hyaluronan synthases (HAS1, 2, and 3), which produce HA polymers with different chain lengths and differ in their rates of synthesis. Whereas high molecular weight HA (> 500 kDa) is anti-angiogenic, anti-inflammatory, and immunosuppressive, low molecular weight HA (10-500 kDa) is highly angiogenic and pro-inflammatory. Functionally, hyaluronan molecules are important for the maintenance of a highly hydrated extracellular matrix in tissues, which is involved in cell adhesion and tissue fibrogenesis. Thus, hyaluronan is a potential biomarker for fibrosis detection.
PRINCIPLE OF THE ASSAY
This assay is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA). The microtiter plate is pre-coated with a polyclonal antibody specific for human hyaluronan. Standards and samples are pipetted into the wells and any human hyaluronan present is bound by the immobilized antibody. After washing away any unbound substances, a biotin labelled polyclonal antibody specific for human hyaluronan is added to the wells. After wash step to remove any unbound reagents, streptavidin-horseradish peroxidase conjugate (STP-HRP) is added. After the last wash step, an HRP substrate solution is added and color develops in proportion to the amount of human hyaluronan bound initially. The assay is stopped, and the optical density of the wells is determined using a microplate reader. Since the increases in absorbance are directly proportional to the amount of captured human hyaluronan, the unknown sample concentration can be interpolated from a reference curve included in each assay.
The lowest level of human hyaluronan that can be detected by this assay is 0.12ng/mL.
Intra-assay Precision (Precision within an assay) C.V. <3.6%.
Inter-assay Precision (Precision between assays) C.V. <5.8%.
PUBLICATIONS CITING THIS PRODUCT
- Fraser JR, et al. (1997). Hyaluronan: its nature, distribution, functions and turnover.
- Ralf Lichtinghagen, et al. (2012) The Enhanced Liver Fibrosis (ELF) score: Normal values, influence factors and proposed cut-off values.