>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
<1.0 EU per 1 μg of the protein by the LAL method.
113 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Substrate: Chondroitin sulfate (Sigma, Catalog # C6737), 10 mg/mL stock in deionized water
96 well UV Plate (Costar, Catalog # 3635)
Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
Dilute rP. vulgaris Chondroitinase ABC to 1 ng/μL in Assay Buffer.
Dilute Substrate to 4 mg/mL in Assay Buffer.
Load 50 μL of 1 ng/μL rP. vulgaris Chondroitinase ABC into the plate, and start the reaction by adding 50 μL of 4 mg/mL Substrate. Include a Substrate Blank containing 50 μL of Assay Buffer and 50 μL of 4 mg/mL Substrate.
Read at an absorbance of 232 nm in kinetic mode for 5 minutes.
Calculate specific activity:
Specific Activity (pmol/min/µg) =
Adjusted Vmax* (OD/min) x well volume (L) x 1012 pmol/mol
ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg)
*Adjusted for Substrate Blank **Using the extinction coefficient 3800 M-1cm-1 ***Using the path correction 0.32 cm Note: the output of many spectrophotometers is in mOD. Per Well:
rP. vulgaris Chondroitinase ABC: 0.050 μg
Chondroitin sulfate: 200 μg
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This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.
Alternate Names for Recombinant P. vulgaris Chondroitinase ABC Protein, CF
Chondroitinase ABC from Proteus vulgaris is a depolymerizing lyase that is especially active on various forms of chondroitin sulfate and dermatan sulfate (1, 2). Chondroitin sulfate contributes a major structural component of cartilage and provides much of the cartilage’s resistance to compression (3). Chondroitin sulfate is also found in the nervous system and at the cell surface, where it plays regulatory roles in development (4, 5) and is involved in pathways related to microbial infection (6). Dermatan sulfate is primarily located in the skin, but is also found in the blood vessels, heart valves, tendons, and lungs, where it plays roles in blood coagulation and wound healing (7). Chondroitinase ABC is also about one hundred-fold less active on hyaluronan, a nonsulfated glycosaminoglycan distributed widely throughout connective, epithelial, and neural tissues (8). Chondroitinase ABC can be used to study the structures and functions of these polysaccharides. For example, in one study it was found to promote axon growth following spinal cord injury, presumably through the degradation of chondroitin sulfate in close proximity to the axons (9). Chondroitinase ABC works through beta -elimination of the 1,4-hexosaminidic bond on the polysaccharides; thereby generates unsaturated disaccharides and tetrasaccharides that exhibit strong UV absorbance at 232 nm (10).
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Publications for Chondroitinase ABC (6877-GH)(2)
We have publications tested in 1 confirmed species: Human.
We have publications tested in 1 application: Bioassay.
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