Aggrecan (ACAN) (Interglobular Domain T 331 - G 458) Protein

Name: Aggrecan (ACAN)

Synonyms: AGC1, SEDK, AGCAN, CSPG1, MSK16, CSPGCP, Agc, cmd, Agc1, Cspg1, ACAN

Epitope: Interglobular Domain T 331 - G 458

Origin (Gene): Human

Application: Blocking Antibody (Inhibition), ELISA

Catalog no.: ABIN368666

Additional Information

Characteristics: Aggrecan Recombinant Interglobular Domain T 331 - G 458 (Aggrecan-IGD1)

Description: Molecular form: The polypeptide connecting human aggrecan globular domains 1 and 2 (T 331 - G 458 ) is expressed in E. coli with a C-terminal His-tag. The recombinant protein contains cleavage sites for aggrecanases (E 373 - A 374 in human aggrecan) and matrix metalloproteinases (N 341 - F 342 in human aggrecan). 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 M r 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 CaCl 2 .

Application Details

Application Notes: Aggrecan interglobular domain 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 M r 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 neoepitop 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 neoepitop 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.

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 (Fig. 1).

Storage: Aggrecan interglobular domain is stable until the expiry date given on the label when stored at -70°C. The protein can be kept at -20°C for several weeks and on ice for several days. Repeated freezing and thawing should be avoided.

Restrictions: For Research Use only

Images

Aggrecan (ACAN) (Interglobular Domain T 331 - G 458) protein