Recombinant P. vulgaris Chondroitinase ABC Protein, CF

Catalog # Availability Size / Price Qty
6877-GH-020
R&D Systems Recombinant Proteins and Enzymes
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Citations (3)
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Recombinant P. vulgaris Chondroitinase ABC Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to hydrolyze chondroitin sulfate. The specific activity is >15,000 pmol/min/μg, as measured under the described conditions.
Source
E. coli-derived p. vulgaris Chondroitinase ABC protein
Ala25-Pro1021, with an N-terminal Met and 6-His tag
Accession #
N-terminal Sequence
Analysis
Met
Predicted Molecular Mass
113 kDa
SDS-PAGE
85-100 kDa, reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

6877-GH

Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Brij-35.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • Assay Buffer: 100 mM Tris, 50 mM NaCl, 10 mM MgCl2, pH 7.5
  • Recombinant P. vulgaris Chondroitinase ABC (rP. vulgaris Chondroitinase ABC) (Catalog # 6877-GH)
  • 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

  1. Dilute rP. vulgaris Chondroitinase ABC to 1 ng/μL in Assay Buffer.
  2. Dilute Substrate to 4 mg/mL in Assay Buffer.
  3. 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.
  4. Read at an absorbance of 232 nm in kinetic mode for 5 minutes.
  5. 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
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Chondroitinase ABC

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).

References
  1. Oike, Y. et al. (1980) Biochem. J. 191:193.
  2. Prabhakar, V. et al. (2009), J. Biol. Chem. 284:974.
  3. Plaas, A.H.K et al. (1998), J. Biol. Chem. 273:12642.
  4. Brittis, P.A. et al. (1992), Science 255:733.
  5. Rauch, W. (1991) J. Biol. Chem. 266:14785.
  6. Fried, M. and Duffy, P.E. Science 272:1502.
  7. Trowbridge, JM and Gallo, RL (2002) Glycobiology 12:117R.
  8. Hamai, A. et al. (1997), J. Biol. Chem. 272:9123.
  9. Garcia-Alias, G. et al. (2009) Nat. Neurosci. 12:1145.
  10. Huang, W. et al. (2007) J. Mol. Biol. 328:623.
Entrez Gene IDs
2827933 (P. vulgaris)
Alternate Names
Chondroitinase ABC

Product Specific Notices

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

Citations for Recombinant P. vulgaris Chondroitinase ABC Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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  1. COVID-19 generates hyaluronan fragments that directly induce endothelial barrier dysfunction
    Authors: KA Queisser, RA Mellema, EA Middleton, I Portier, BK Manne, F Denorme, EJ Beswick, MT Rondina, RA Campbell, AC Petrey
    JCI Insight, 2021;0(0):.
    Species: Human
    Sample Types: Plasma
    Applications: Bioassay
  2. Neurocan, an extracellular chondroitin sulfate proteoglycan, stimulates neuroblastoma cells to promote malignant phenotypes
    Authors: Z Su, S Kishida, S Tsubota, K Sakamoto, D Cao, S Kiyonari, M Ohira, T Kamijo, A Narita, Y Xu, Y Takahashi, K Kadomatsu
    Oncotarget, 2017;8(63):106296-106310.
    Species: Human
    Sample Types: Cell Culture Supernates
    Applications: Bioassay
  3. Heparin-binding protein is important for vascular leak in sepsis
    Authors: Adam Linder
    Intensive Care Med Exp, 2016;4(1):33.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay

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