Cat# H1000 and H2000
The Biocolor Purple-Jelley Hyaluronan Assay is a quantitative assay used to measure the quantity of hyaluronic acid in mammalian tissue.
It is a dye-based method with a detection limit of 0.2μg. The dye used is ‘Stains-all’.
The assay manual includes methods for the user to remove tissue protein and sulfated glycosaminoglycans before measuring the isolated hyaluronic acid. Also included in the manual is an outline guide to the distribution of hyaluronic acid present in various animal tissues – heart, kidney, lung, leg muscle and skin. All values were obtained using this assay. In adult animals the hyaluronic acid percentage gradually increases during ageing as muscle and fat mass decrease.
About Hyluronic Acid
Hyaluronic acid (HA), also known as hyaluronan, is a non-sulphated glycosaminoglycan (GAG) and is composed of repeating polymeric disaccharides of D-glucuronic acid and N-acetyl-D-glucosamine linked by a glucuronidic β (1→3) bond. In aqueous solutions HA forms specific stable tertiary structures. Despite the simplicity in its composition, without variations in its sugar composition or without branching points, HA has a variety of physicochemical properties. HA polymers occur in a vast number of configurations and shapes, depending on their size, salt concentration, pH, and associated cations. Unlike other GAG, HA is not covalently attached to a protein core, but it may form aggregates with proteoglycans. HA encompasses a large volume of water giving solutions high viscosity, even at low concentrations.
HA is widely distributed, from prokaryotic, to eukaryotic cells. In humans, HA is most abundant in the skin, (accounting for 50% of the total body HA), the vitreous of the eye, the umbilical cord, and synovial fluid, but it is also present in all tissues and fluids of the body, such as skeletal tissues, heart valves, the lung, the aorta, the prostate, tunica albuginea, corpora cavernosa and corpus spongiosum of the penis. HA is produced primarily by mesenchymal cells but also by other cell types.
Considerable evidence exists regarding the functional role of HA in molecular mechanisms and, consequently, the potential role of HA for the development of novel therapeutic strategies for many diseases.
Functions of HA include the following: hydration, lubrication of joints, a space filling capacity, and the framework through which cells migrate. The synthesis of HA increases during tissue injury and wound healing and HA regulates several aspects of tissue repair, including activation of inflammatory cells to enhance immune response and the response to injury of fibroblasts and epithelial cells. HA also provides the framework for blood vessel formation and fibroblast migration, that may be involved in tumor progression. The correlation of HA levels on the cell surface of cancer cells with the aggressiveness of tumors has also been reported.
The size of HA appears to be of critical importance for its various functions described above. HA of high molecular size, usually in excess of 1,000 kDa, is present in intact tissues and is antiangiogenic and immunosuppressive, whereas smaller polymers of HA are distress signals and potent inducers of inflammation and angiogenesis.
Tissue Extracts. Removal of tissue protein before assay is essential in order to isolate the HA from potentially dye binding proteins; and sulfated glycosaminoglycans need to be removed before measuring the isolated HA, from a two step critical electrolyte salting out process (CEC). Methods for protein removal and sGAG removal can be found in the HA manual.
Examples of Results
Distribution of HA in Tissue
Species Tissue Concentration (µg/g) Mouse heart 48 kidney 11 lung 14 leg muscle 20 skin 307 Rat heart 83 kidney 28 lung 17 leg muscle 67 skin 510 Rabbit heart 138 kidney 27 lung 45 leg muscle 50 skin 428
Hyaluronic acid, Hyaluronan, HA, Purple-Jelly Hyaluronan Assay
In the links below you will find the Purple-Jelley Hyaluronan Assay protocol, publications that reference the Purple-Jelley Hyaluronan Assay, and the Purple-Jelley Hyaluronan Assay MSDS.
Want to learn more about Hyaluronan? We have compiled the links below which contain information that you may find interesting.
- Hyaluronic acid as a bioactive component for bone tissue regeneration: Fabrication, modification, properties, and biological functions
- The application of hyaluronic acid in bone regeneration
- Hyaluronic Acid-Based Hydrogels: from a Natural Polysaccharide to Complex Networks
Ilex Life Sciences LLC is an official distributor of Biocolor products.