Glycosaminoglycan Detection Kits
| Product | Catalog # | Price (USD) | |
|---|---|---|---|
 |
Glycosaminoglycans Assay Kit | 6022 | 321.00 |
 |
Hyaluronan Competition Assay Kit | 6048 | 389.00 |
 |
Hyaluronan Sandwich Assay Kit | 6049 | 389.00 |
Reagents for Sample Preparation
Sulfated Glycosaminoglycans
Glycosaminoglycans (GAGs) are negatively charged polysaccharides located in most connective tissues and extracellular matrix (ECM), as well as on the surfaces of many cell types. Consisting of repeating core disaccharide units, GAGs are categorized into four types: heparin/heparan sulfate, chondroitin/dermatan sulfate, keratin sulfate and non-sulfated hyaluronan. Sulfated GAGs in the ECM exist as proteoglycan which typically consists of multiple glycosaminoglycan chains attached to a core protein. As a highly organized ECM, articular cartilage is composed of type II collagen, hyaluronan, link protein, and chondroitin sulfate-rich proteoglycans, which provide the osmotic resistance necessary for cartilage to resist compressive loads. Chondrex, Inc. provides a sulfated GAGs assay kit using the cationic dye, 1,9 dimethylmethylene blue (DMB) which binds to highly charged sulfated GAGs, not including hyaluronan. This kit utilizes an improved DMB solution minimizing interference with negatively charged contaminants, such as DNA and RNA, and chondroitin sulfate as an appropriate standard for the analysis of ECM in cartilage.
Table 1 provides a useful matrix to determine the ideal solubilization protocol for the analyte of interest. In most cases, two different protocols will need to be used. For example, protocol A extracts GAGs, but fails to completely extract collagen and DNA, resulting in underestimated values (1,2). Protocol B solubilizes GAGs (3) and DNA (4), but degrades collagen. Finally, protocol C (Chondrex, Inc.'s protocol) can be used to measure GAGs and collagen (5), but not DNA.
Hyaluronans
Hyaluronan (HA: also called Hyaluronic acid) is a non-sulfated, unbranched glycosaminoglycan (GAG) composed of repeating disaccharide units of D-glucuronic acid and N-acetyl-D-glucosamine (6). Nearly all vertebrate cells can produce HA, with its expression closely linked to processes such as tissue expansion and cell movement (7). HA is primarily found in the extracellular and pericellular matrices, though it can also be present intracellularly. Its key biological roles include maintaining the elastoviscosity of fluid connective tissues such as joint synovial fluid and the vitreous humor in the eye, regulating tissue hydration and water transport, and organizing proteoglycans in the extracellular matrix. Additionally, HA is involved in receptor-mediated processes like cell detachment, mitosis, migration, tumor progression and metastasis, and inflammation. It also plays a vital role in binding water and lubricating movable parts of the body, such as joints and muscles (8). HA receptors on effector cells interact with HA and are involved in HA-related cell behavior and uptake (9).
References
- C. D. Hoemann, J. Sun, V. Chrzanowski, M. D. Buschmann, A multivalent assay to detect glycosaminoglycan, protein, collagen, RNA, and DNA content in milligram samples of cartilage or hydrogel-based repair cartilage. Anal Biochem 300, 1-10 (2002).
- S. W. Sajdera, V. C. Hascall, Proteinpolysaccharide complex from bovine nasal cartilage. A comparison of low and high shear extraction procedures. J Biol Chem 244, 77-87 (1969).
- R. W. Farndale, D. J. Buttle, A. J. Barrett, Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. Biochem Biophys Acta 883, 173-177 (1986).
- Y. J. Kim, R. L. Sah, J. Y. Doong, A. J. Grodzinsky, Fluorometric assay of DNA in cartilage explants using Hoechst 33258. Anal Biochem 174, 168-176 (1988).
- K. von der Mark, M. van Menxel, H. Wiedemann, Isolation and characterization of new collagens from chick cartilage. Eur J Biochem 124, 57-62 (1982).
- I. Hargittai, M. Hargittai. Molecular structure of hyaluronan: an introduction. Struct Chem 19, 697-717 (2008).
- V. Hascal, J. Esko. “Hyaluronan” in Essentials of Glycobiology (Cold Spring Harbor Laboratory Press, ed. 3, 2017).
- J. Necas, L. Bartosikova, P. Brauner, J. Kolar. Hyaluronic acid (hyaluronan): a review. Vet Med (Praha) 53(8), 397-411 (2008).
- B. Toole. Hyaluronan and its binding proteins, the hyaladherins. Curr Opin Cell Biol 2(5), 839-44 (1990).