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  • Brugia malayi chitinase (BmCHT1)

    Discontinued Date

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

    Description

    Brugia malayi chitinase (BmCHT1) is expressed in the microfilarial stage, the first larval stage, of the organism and is thought to be important in the exsheathment process of the microfilaria (1). Exsheathment is required for further development of the microfilaria once ingested by the mosquito vector (2). The microfiliaria of Brugia malayi have been shown to have chitin in their sheaths (3). Antisera to the chitinase temporarily cleared the microfilaria from the bloodstream of infected jirds (1,4).

    Product Source

    Isolated from the supernatant of Spodoptera frugiperda (Sf9) insect cells infected with an AcNPV chiA minus recombinant baculovirus carrying the BmCHT1 gene cloned from Brugia malayi (kindly provided by J. Fuhrman) (1).

    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 1 minute at 25°C in a total reaction volume of 100 μl.

    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: 54 kDa

    References

    1. Fuhrman, J.A. et al. (1992). Proc. Natl. Acad. Sci. USA. 89, 1548-1552.
    2. Fuhrman, J.A. (1994). Exp. Parasitol. 79, 85-88.
    3. Fuhrman, J.A. and Piessens, W.F. (1985). Mol. Biochem. Parasitol. 17, 93-104.

    Tech Tips

    Enzyme is active from room temperature to 37°C.

    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.

    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.

    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.