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  • Onchocerca volvulus chitinase (OvCHT1)

    Discontinued Date

    01/01/2013
    cloned at neb recombinant incubation temp heat inactivation
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    Categories:
    Discontinued Products

    Description

    Onchocerca volvulus chitinase (OvCHT1) is expressed during development in the black fly vector in the infective L3 larvae and then secreted during post-infective development in the human host (1, 2). The specific role of the chitinase is not yet clear.

    Product Source

    Isolated from the supernatant of Spodoptera frugiperda (Sf9) insect cells infected with an AcNPV chiA minus recombinant baculovirus carrying the OvCHT1 gene cloned from Onchocerca volvulus.

    Properties and Usage

    Unit Definition

    One unit is defined as the amount of enzyme required to release the equivalent fluorescence produced by 1 pmol 4-methylumbelliferone from the substrate 4-methylumbelliferyl-N,N',N"-triacetyl-β-chitotrioside in a total reaction volume of 100 μl in 1 minute at 25°C in 200 mM NaCl with 20 mM NaPO4 (pH 6.0), 1 mM EDTA and 20 µM 4-methylumbelliferyl-N,N',N"-triacetyl-β-chitotrioside.

    Storage Temperature

    -20°C

    Storage Conditions

    20 mM Tris-HCl
    200 mM NaCl
    1 mM EDTA
    50% Glycerol
    0.1% Triton® X-100
    pH 7.5 @ 25°C

    Heat Inactivation

    65°C for 20 min

    Molecular Weights

    Apparent: 65 kDa
    Theoretical: 53 kDa

    Quality Control

    Quality Control Assays

    The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
    • Functional Test (Chitin Binding):
      The Chitinase is tested by incubation with chitin resin in binding buffer. After multiple washes no chitinase is detected in the unbound or wash samples as determined by measurement of activity or by analysis on SDS-PAGE
    • Functional Test (Chitin Radioactivity Release):
      The Chitinase is tested in a reaction with tritiated Chitin. After incubation the percentage of released soluble cpm is determined at various time points up to 24 hours.

    References

    1. Wu, Y. et al. (2001). J. Biol. Chem.. 276, 42557-42564.
    2. Wu, Y. et al. (1996). Mol. Biochem. Parasitol.. 75, 207-219.

    Supporting Documents

    Material Safety Datasheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.

    Datacards

    The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.
    Enzyme is active from room temperature to 37°C.