Catalog Number:
5028003-B
Aggrecan IGD (interglobular domain) recombinant protein is used as substrate for aggrecanases and matrix metalloproteinases.
Overview
Overview:
Aggrecan IGD recombinant protein is used as substrate for aggrecanases and matrix metalloproteinases. For proteinase activity measurements, the protein is incubated with proteinase for various time intervals. Thereafter, aliquots of the incubation mixture are analyzed by SDS-PAGE or by ELISA. Upon cleavage with aggrecanases, the apparent Mr of aggrecan interglobular domain in SDS-PAGE is reduced from 21 kDa to about 13 kDa (Fig. 1). Quantitative measurement of aggrecanase cleavage requires a neoepitope antibody with specificity for the N-terminus A R G S V I L T … appearing upon hydrolysis. The fragment with the newly formed N-terminus is fixed by the neoepitope antibody to a microplate and quantified with an anti-His-tag antibody. In analogy, cleavage by matrix metalloproteinases can be measured with antibodies to neoepitopes appearing upon action of these enzymes.
Related Symbols/Terms:
- Aggrecan 1
- Chondroitin sulfate proteoglycan 1
- Cartilage-specific proteoglycan core protein
- AGC1
- CSPG1
- ACAN Gene
- Recombinant aggrecan IGD protein
Data/Specifications
Data/Specifications:
Molecular Form:
The polypeptide connecting human aggrecan globular domains 1 and 2 (T331 - G458) is expressed in E. coli with a C-terminal His-tag. The recombinant protein contains cleavage sites for aggrecanases (E373 - A374) and matrix metalloproteinases (N341 - F342). It comprises the following amino acids:
T A E D F V D I P E N F F G V G G E E D I T V Q T V T W P D M E L P L P R N I T E
G E A R G S V I L T V K P I F E V S P S P L E P E E P F T F A P E I G A T A F A E V
E N E T G E A T R P W G F P T P G L G P A T A F T S E D L V V Q V T A V P G Q P
H L P G G (His-tag)
Main cleavage sites are indicated by arrows. The calculated Mr of the His-tagged protein is
15,493 Da. The protein is solubilized in 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 5 mM CaCl2.
Purity:
The recombinant aggrecan interglobular domain appears as a major band at about 21 kDa in SDS-PAGE. It represents more than 90% of total protein in the preparation.
Storage:
Aggrecan interglobular domain is stable until the expiry date given on the label when stored at -70 °C. The enzyme can be kept at -20 °C for several days. Repeated freezing and thawing should be avoided.
Size: 500 ug
Source: E. coli