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CS1030

Sigma-Aldrich

Chitinase Assay Kit, Fluorimetric

sufficient for 200 multiwell tests

Synonym(s):

Chitinase Detection Kit

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About This Item

UNSPSC Code:
12161503
NACRES:
NA.84

usage

sufficient for 200 multiwell tests

Quality Level

shipped in

wet ice

storage temp.

−20°C

Gene Information

human ... CHIT1(1118)

General description

The kit assay is based on the enzymatic hydrolysis of chitinase substrates. This enzymatic hydrolysis releases 4-methylumbelliferone (4MU), which upon ionization in basic pH, can be measured fluorimetrically at an excitation wavelength of 360 nm and an emission wavelength of 450 nm. The use of fluorimetric substrates provides a very sensitive detection system.

Application

The Chitinase Assay Kit provides all the reagents required for efficient and sensitive detection of chitinase activity in fungal and bacterial growth media, macrophage lysates, and purified enzyme preparations. In addition, the kit provides three different substrates for the detection of the various types of the chitinolytic activity:
  • 4-Methylumbelliferyl N,N′-diacetyl-β-D-chitobioside - substrate suitable for exochitinase activity detection (chitobiosidase activity)
  • 4-Methylumbelliferyl N-acetyl-β-D-glucosaminide - substrate suitable for exochitinase activity detection (β-N-acetylglucosaminidase activity)
  • 4-Methylumbelliferyl β-D-N,N′,N′′-triacetylchitotriose - substrate suitable for endochitinase activity detection

Biochem/physiol Actions

Chitinase catalyzes the hydrolytic cleavage of the β-1→4-glycoside bond present in biopolymers of N-acetylglucosamine, primarily in chitin. Chitinases are widely distributed in living organisms and are found in fungi, bacteria, parasites, plants, and animals. They are classified in families based on amino acid sequence similarities.
The chitinolytic enzymes are also categorized based on their enzymatic action on chitin substrates. Endochitinases are defined as the enzymes catalyzing the random cleavage at internal points in the chitin chain. Exochitinases catalyze the progressive release of acetylchitobiose or N-acetylglucosamine from the non-reducing end of chitin, and are referred to as chitobiosidase and β-N-acetylglucosaminidase, respectively.
Chitinases perform different functions in different organisms. In bacteria, they are mainly involved in nutritional processes. In yeast and various fungi, these enzymes participate in morphogenesis. In animals and plants, chitinases primarily play a role in the defense of the organism against pathogen attack.

Suitability

The kit was tested and found suitable for Trichoderma viride and Streptomyces griseus, along with Hela, Jurkat, CHO, NIH-3T3, U-837 mammalian cell lines, human macropages, rat lung, kidney, liver, and brain tissue.

Preparation Note

Use ultrapure water for preparation of reagents.

Kit Components Only

Product No.
Description

  • Assay Buffer 25 mL

  • 4-Methylumbelliferyl N-acetyl-β-D-glucosaminide 5 mg

  • 4-Methylumbelliferyl β-D-N,N′-diacetylchitobioside hydrate 5 mg

  • 4-Methylumbelliferyl β-D-N,N′,N′′-triacetylchitotriose 5 mg

  • Chitinase from Trichoderma viride 1 mg

  • 4-Methylumbelliferone Standard Solution, 50 mg/mL 1 mL

  • Sodium Carbonate 2 g

  • Dimethyl Sulfoxide 1 mL

See All (8)

Pictograms

Health hazard

Signal Word

Danger

Hazard Statements

Hazard Classifications

Eye Irrit. 2 - Resp. Sens. 1

Storage Class Code

10 - Combustible liquids

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

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Computational modeling and functional characterization of a GgChi: A class III chitinase from corms of Gladiolus grandiflorus
Rafiq M, et al.
The Kaohsiung Journal of Medical Sciences (2018)
S S Thimoteo et al.
Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas, 50(1), e5658-e5658 (2017-01-12)
Chitinases are hydrolases that degrade chitin, a polymer of N-acetylglucosamine linked β(1-4) present in the exoskeleton of crustaceans, insects, nematodes and fungal cell walls. A metagenome fosmid library from a wastewater-contaminated soil was functionally screened for chitinase activity leading to
Daniel Hartmann et al.
Future oncology (London, England), 11(2), 193-203 (2014-07-22)
N-acetyl-glucosaminidase (NAG) is a potential marker of genotoxicity. We retrospectively analyzed plasma NAG and clinico-pathologic features in advanced gastrointestinal adenocarcinoma patients. Plasma from 118 patients and 51 healthy volunteers was analyzed for associations between NAG levels and age, disease presence
Shinichiro Enomoto et al.
Cell host & microbe, 21(5), 629-636 (2017-05-12)
Sodalis praecaptivus is a close relative and putative environmental progenitor of the widely distributed, insect-associated, Sodalis-allied symbionts. Here we show that mutant strains of S. praecaptivus that lack genetic components of a quorum-sensing (QS) apparatus have a rapid and potent killing
Jéssica Rodrigues et al.
Scientific reports, 5, 11717-11717 (2015-07-15)
Cryptococcus gattii is one of the causative agents of human cryptococcosis. Highly virulent strains of serotype B C. gattii have been studied in detail, but little information is available on the pathogenic properties of serotype C isolates. In this study

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