| 产品目录 |
|
|
 |
|
|
 |
| 硫酸软骨素酶ABC/Chondroitinase ABC |
|
- CATALOG # 100330-1A
- Package Size: 10 units
- Chondroitinase ABC
- (from Proteus vulgaris)
- EC 4.2.2.4
- SYSTEMATIC NAME:
- Chondroitin ABC lyase
- TRIVIAL NAME:
- Chondroitin ABC eliminase
-
- Chondroitinase ABC purified from Proteus vulgaris by precipitation with ammonium sulfate followed by DEAE-Cellulose chromatography catalyzes the eliminative cleavage of N-acetylhexosaminide linkages in chondroitin 4-sulfate, chondroitin 6-sulfate, dermatan sulfate, chondroitin, and hyaluronic acid, yielding mainly disaccharides with D(4) - hexuronate at the non-reducing ends.
-
- This enzyme does not act on keratan sulfate, heparin, and heparan sulfate. The initial rates of enzymatic degradation of chondroitin sulfate C, dermatan sulfate, chondroitin and hyaluronic acid were 1.0, 0.4, 0.2, and 0.02, respectively, relative to the rate of chondroitin sulfate A degradation.
-
- The enzyme can be used for selective removal of the chondroitin sulfate or dermatan sulfate side chains from proteoglycans, yielding a protein - enriched core molecule(4).
- SPECIFICATIONS:
-
| Appearance: |
Lyophilized powder
Notice that bovine serum albumin (less than 1mg/vial) is added as a stabilizer. |
| Activity: |
Not less than 5units/vial |
| Contaminants: |
Chondro-4-sulfatase: less than 5.0 x 10-3units/vial
Chondro-6-sulfatase: less than 2.3 x 10-5units/vial
(as measured by increase (or decrease) of A585 after Morgan-Elson reaction) |
| Optimum pH: |
8.0 (for chondroitin sulfate)
6.2 (for hyaluronic acid) |
| Recommended reaction temperature: |
37oC |
| Molecular weight: |
120,000-145,000 (gel filtration) |
| Activators: |
Acetate (0.05M) |
| Inhibitors: |
Zn2+ (10-3M ZnCl2 inhibits by 100%)
Heparin (an equimolar amount of heparin inhibits by ca. 70%) |
- ASSAY FOR ENZYME ACTIVITY:
-
- Unit Definition
- One unit is defined as the quantity of the enzyme that catalyzes the formation of 1 mmole of unsaturated disaccharide from chondroitin 6-sulfate per minute at 37oC, pH 8.0(1).
-
- Method
- The assay is based on that of Yamagata et al.(1). Reaction mixture
| Substrate and buffer solution |
0.1 mmole chondroitin 6 - sulfate in
10 m l of 0.4M Tris - HCl buffer (pH 8.0)
10 ml of 0.4M CH3COONa
10 ul of 0.1% bovine serum albumin (BSA)
70 ul of distilled water |
| Enzyme solution |
20 ml of suitably diluted enzyme (1-5milli units);
0.1% BSA is used for the dilution of enzyme. |
|
| Total volume |
120 ul...
-
|
| Reaction |
Incubated at 37oC for 10 minutes. Stopped by boiling for 1minute. |
| Morgan - Elson reaction... |
To enzyme reaction mixture (120 ul) add 0.1ml of 5% K2B4O7 (pH 9.0), and heat in a boiling water bath for 7 minutes.After cooling, add 1ml of glacial acetic acid, mix, add 0.4ml of the Morgan - Elson reagent, and incubate at 37oC for 20minutes.
Measure A585. |
- Calculation
Enzyme unit (U/ml) = |
A585 |
x |
0.1 |
x |
1 |
x |
1 |
|
|
|
|
| 2.38 |
G |
10 |
E |
- G: Adsorption of 0.1 mmole GalNAc (A585)
- E: Volume of enzyme solution (ml)
- STORAGE:
- The enzyme is stable for two years when stored at - 20oC. The solution (1unit/100 ul) is stable for one week when stored at 4oC.
-
- REFERENCES:
- 1) Yamagata, T., saito, H., Habuchi, O. and Suzuki, S.: J. Biol. Chem., 243, 1523 (1968)
- 2) Saito, H., Yamagata, T. and Suzuki, S.: J. Biol. Chem., 243, 1536 (1968)
- 3) Suzuki, S., Sato, H., Yamagata, T., Anno, K., Seno, N., Kawai,k Y. and Furuhashi, T.: J. Biol. Chem., 243, 1543 (1968)
- 4) Oike, Y., Kimata, K., Shinomura, T., Suzuki, S., Takahashi, N. and Tanabe, K.: J. Biol. Chem., 257, 9751 (1982)
- NOTE:
- For laboratory use only. Not for drug, household or other uses.
|
 |
|
|
