Onchocerca volvulus chitinase (OvCHT1)

  • This product was discontinued on 01/01/2013
cloned at neb recombinant incubation temp heat inactivation
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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

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.

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.

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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. Further information regarding NEB product quality can be found here.
  • 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.

Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

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.