Glycosaminoglycan ELISA Kit (GAGs)

Full Name: Glycosaminoglycan ELISA Kit (GAGs)
Reactivity: Human
Sample Type: Serum, Biological Fluids, Tissue Homogenates, Plasma
Sensitivity: 0.938 ng/ml

INTRODUCTION

A complex component of glycans, glycosaminoglycan (GAGs) are a complex family of highly sulfated, polydisperse linear polysaccharides. There are four categories that are essentially based on the differing disaccharide repeat units; keratan sulfate, chrondroitin sulfate/dermatan, heparin/heparan sulfate and hyaluronan. Keratan sulfate is the largest and most abundant GAG in the body. It is a sulfated version of glucosamine and it forms a scaffold for the extracellular matrix by binding to collagen, reticular fibers, elastic fibers, etc., in addition to proteoglycan parts of cells that are exposed on cell membranes. Keratan sulfate chains can be easily broken apart into smaller segments called disaccharides with only two different types: Glucuronic acid and iduronic acid. Chondroitin sulfates/dermatan are mostly found in cartilage and tendons.

Glycosaminoglycan activities are usually initiated through the interactions of many different types of proteins. One of the main functions is within the ECM is in the regulation of mechanical activities in a tissue this can include such things as; cell adhesion, immune cell function, cell proliferation, formation of collagen structure and growth factor signalling. The structure and composition of the ECM is determined by two main proteins; collagen and proteoglycan.

GAGs have a unique polymer structure that are composed mainly of repeating disaccharide units linked by alpha-1,3 bonds. The creation of a compound between hexosamines and a fatty acid to produce a lipid raft domain is the initial step in the biosynthesis of proteoglycans. This domain is then cleaved into two domains, A and B. Domain A contains mannosyl-glycans while domain B contains glucosyl-glycans. Glycoproteins fold into an alpha helical structure to give rise to proteoglycan core protein with multiple globular appendages called glycophosphatidic acid chains which are stabilized by hemagglutinin, galactose and mannose residues.

INTENDED USE

Human glycosaminoglycan ELISA kit is designed for analysing concentrations of GAGs (glycosaminoglycan) present in human samples of tissue homogenates, serum, plasma and other biological fluids.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

One 96-Well Plate: Pre-coated with anti GAGs antibody.
Standards: Lyophilized recombinant.
Sample/Standard Dilution Buffer.
Biotinylated-labelled Antibody.
Antibody Dilution Buffer.
HRP-Streptavidin Conjugate (SABC).
SABC Dilution Buffer.
TMB Substrate.
Wash Buffer (25x).
Plate Sealer.
Product Instructions.

TYPICAL RESULTS

For this glycosaminoglycan ELISA kit it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 1.562, 3.125, 6.25, 12.5, 25, 50, 100 ng/ml.
Reactivity: Human
Sensitivity: 0.938 ng/ml
Range: 1.563 – 100 ng/ml
Principle: Competitive
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for GAGs, no cross reactivity or interference between GAGs and analogues was detected.
– Recovery: Serum (98 – 103%), EDTA Plasma (91 – 101%), Heparin Plasma (87 – 103%).
– Linearity: Serum (83 – 94%), EDTA Plasma (89 – 100%), Heparin Plasma (83 – 97%).
– Precison Intra-Assay: CV < 8%.
– Precison Inter-Assay: CV < 10%.
– Stability: Less than 10%.

REFERENCES

Collagen/glycosaminoglycan-based matrices for controlling skin cell responses. Biol Chem. (2021) 402 (11): 1325-1335. Anderegg U., et al.
Chemical Modification of Glycosaminoglycan Polysaccharides. Molecules. (2021) 26 (17): 5211. Palhares LCGF., et al.
The effect of glycosaminoglycans (GAGs) on amyloid aggregation and toxicity. Molecules. (2015) 20 (2): 2510-28. Iannuzzi C., et al.
Glycosaminoglycan-Based Cryogels as Scaffolds for Cell Cultivation and Tissue Regeneration. Molecules. (2021) 26 (18): 5597. Wartenberg A., et al.

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