rEGCase I assisted by Activator II

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Synonyms:

• Recombinant Endoglycoceramidase I assisted by Activator II
• Recombinant - Rhodococcus equi EGCase I expressed in Rhodococcus erythropolis
• Recombinant Endoglycoceramidase I with Activator II

Product Documents:

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General Information | Specifications & Properties | Safety & Regulations | Applications & Literature

Supplemental Product Information

Explanatory notes for this page

Size	Unit Price	Belgium	Japan*	Quantity
1VIAL	€358.00	2	7

*Stock available in Belgium will be delivered in 1 to 3 days
*Stock available in Japan will be delivered in 1 to 2 weeks (excludes regulated items and dry ice shipments).

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Product Information

• Quantity : 300 mU per vial (3 mU/μL) and 50 nmol Activator II per vial (0.5 nmol/μL)
• Product Form : Purified recombinant EGCase I and Activator II, Liquid
• Storage buffer : 20 mM Sodium phosphate, 50 mM NaCl buffer, pH 7.0
• Preservative : None
• Stabilizer : None

Description

• Endoglycoceramidase I (EGCase I) is a glycolipid-specific hydrolase that cleaves the glycosidic linkage between oligosaccharide and ceramide of various glycosphingolipids (GSLs) under a detergent-containing condition. On the other hand, Activator II is capable of inducing activity of EGCase without any detergent reagents. This product is provided as a mixture-form of EGCase I and Activator II. By using this product, cell-surface glycosphingolipids of living cells are hydrolyzed without cell disruption caused by addition of detergent. The EGCase I gene is derived from Rhodococcus equi and its recombinant protein is expressed in Rhodococcus erythropolis.

Unit Definition

• One unit (U) of EGCase I is defined as the amount required to hydrolyze 1 μmol of ganglioside GM1 per minute at 37°C under the condition of pH 5.5.

Assay Conditions

• Reaction conditions: 50 mM sodium acetate, pH 5.5
  The reaction mixture includes the enzyme and 100 nmol of GM1 in 50 μL (without detergent). The mixture is incubated at 37°C for 15 minutes. Quantification of reducing end of released oligosaccharides is measured by the modified Somogyi-Nelson method (see the following reference).
• [Reference]
  Determination of Glucose by a Modification of Somogyi-Nelson Method
  C. Hatanaka, Y. Kobara, Agric. Biol. Chem. 1980, 44, 2943.
• 
  
• Regarding hydrolysis of cellular GSLs, see the reference (Methods Mol. Biol.) below.
• [Reference]
  Assays and Utilization of Enzymes Involved in Glycolipid Metabolism in Bacteria and Fungi
  M. Ito, Y. Ishibashi, T. Watanabe, J. Iwaki, T. Kurita, N. Okino, Methods Mol. Biol. 2023, 2613, 229.

Storage

• Store at -20°C. Avoid repetitive freezing and thawing. After thawing the product, it’s recommended to store at 4°C and use it within a few weeks.

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