Inflammatory Assays

Aggrecanase Activity and Sensitive Assays

Aggrecanase Activity ELISA				Catalog # ACT-AGG.96
Format	Sandwich ELISA				View product insert
Size/Volume	96 well
Species	human				Request Quote
Sample Type	-
Sample Preparation	-
Sample Size	5 uL for Aggrecanase Module 100 uL for ELISA Module
Standard Curve Range	0.47 - 30 nM for Aggrecanase Module 0.062 - 4 nM for ELISA Module
Sensitivity	0.025 nM
Assay Length	3.75 hours

OVERVIEW

Specific quantitative assay for the determination of Aggrecanase activity.

 

Sensitive Aggrecanase Activity ELISA				Catalog # SEN-AGG.96
Format	Proteolysis Reaction/Sandwich ELISA				View product insert
Size/Volume	96 well
Species	human				Request Quote
Sample Type	cell culture supernates
Sample Preparation	-
Sample Size	5 uL for Aggrecanase Module 100 uL for ELISA Module
Standard Curve Range	0.062 - 2 nM Aggrecanase Module 0.022 - 1.4 nM ELSIA Module
Sensitivity	2 pM
Assay Length	3.75 hours

OVERVIEW

This assay is intended for the quantitative determination of low aggrecanase activity.

 

Aggrecanase Activity Assay - a sensitive method to measure activities of ADAMTS1, ADAMTS4 and ADAMTS5

Aggrecanases and their functions
Aggrecanases ADAMTS1, ADAMTS4 and ADAMTS5 are members of the ADAMTS-family of proteases (ADAMTS = A disintegrin and metalloprotease with thrombospondin motif). They are  synthesized as multidomain proteins consisting of prodomains, protease, disintegrin, thrombospondin and cys-rich domains followed by spacer regions and additional thrombospondin motifs. The prodomains are cleaved-off intracellularly and the enzymes are processed further from their C-terminal ends after secretion.

Aggrecanases cleave specific peptide bonds in aggrecan, the main proteoglycan of articular cartilage. They hydrolyze also other lecticanes as versican and brevican. Convincing evidence indicates that aggrecanases are mainly responsible for excessive aggrecan degradation in osteoarthritic joints. 1,2

References: 1) Lohmander, L.S. et al. (1993) Arthritis Rheumat. 36, 1214-1222;  2) Malfait, A. M. et al. (2002) J. Biol. Chem. 277, 22201-22208

Principle of the Assay

 

The Aggrecanase Activity Assay measures activity of aggrecanases and is used to screen and characterize aggrecanase inhibitors. This assay consists of two modules, the Aggrecanase Module and the ELISA Module. In the Aggrecanase Module, a recombinant fragment of human aggrecan interglobular domain (aggrecan-IGD) is first digested with aggrecanase. Proteolytic cleavage of the substrate releases an aggrecan peptide with the N-terminal sequence ARGSVIL (ARGSVIL-peptide). The ARGSVIL-peptide is then quantified in the ELISA Module using two monoclonal anti-peptide sequence antibodies.

The Sensitive Aggrecanase Activity assay measures activities of aggrecanase in the pM concentration range. The high sensitivity is acheived with an engineered aggreganase substrate derived from aggrecan interglobular domain.

 

Product inserts are for information use only.