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Merck

CS0980

Chitinase Assay Kit

sufficient for 100 multiwell tests

Synonym(s):

Chitinase Activity Detection Kit

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About This Item

NACRES:
NA.54
UNSPSC Code:
12161503

Product Name

Chitinase Assay Kit, sufficient for 100 multiwell tests

usage

sufficient for 100 multiwell tests

absorbance ratio

405 nm (Absorbance)

shipped in

wet ice

storage temp.

2-8°C

Quality Level

200

Gene Information

human ... CHIA(27159), CHIT1(1118)

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Analysis Note

The kit assay is based on the enzymatic hydrolysis of chitinase substrates. This hydrolysis releases p-nitrophenol (4-nitrophenol), which upon ionization in basic pH can be measured colorimetrically at 405 nm.

Application

Chitinase Assay Kit has been used in screening chitinase activity of recombinant chitinase from yeast, Metschnikowia fructicola, bacterial endophyte 3A12 and Clostridium difficile spores.

The kit provides all the reagents required for efficient 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-Nitrophenyl N,N′-diacetyl-β-D-chitobioside - substrate suitable for exochitinase activity detection (chitobiosidase activity)
  • 4-Nitrophenyl N-acetyl-β-D-glucosaminide - substrate suitable for exochitinase activity detection (β-N-acetylglucosaminidase activity)
4-Nitrophenyl 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.

General description

The Chitinase Assay Kit is based on the enzymatic hydrolysis of chitinase substrates. The hydrolysis releases p-nitrophenol, which can be measured colormetrically at 405 nm.

pictograms

Health hazard
GHS08

signalword

Danger

Hazard Classifications

Carc. 2 - Eye Irrit. 2 - Resp. Sens. 1 - STOT RE 2 Oral

target_organs

Liver,Kidney

Storage Class

10 - Combustible liquids

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Certificates of Analysis (COA)

Lot/Batch Number
0000503497
0000503497
0000335990
0000335990
0000268892

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  1. Which document(s) contains shelf-life or expiration date information for a given product?

    If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.

  2. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  3. How do I find price and availability?

    There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers.

  4. What is the Department of Transportation shipping information for this product?

    Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product. 

  5. What has Product CS0980, Chitinase Assay Kit, been tested on?  

    This kit has been tested on Trichoderma viride, Streptomyces griseus and human macrophages.

  6. What is the recommended substrate concentration for Product CS0980, Chitinase Assay Kit?  

    A 1mg/mL substrate solution is recommended. If required, the 4-Nitrophenyl-beta-D-N,N',N''-triacetylchitotriose and the 4-Nitrophenyl N,N'-diacetyl-beta-D-chitobioside can be used at lower concentrations in the reaction (0.5 or 0.2mg/mL respectively).

  7. How do you make the substrate in Product CS0980, Chitinase Assay Kit, go into solution?

    The substrates do not dissolve easily in the buffer.  It may take ~1 hour of shaking to completely dissolve the substrates. Use of a larger container (15 mL) may aid dissolution.

  8. What is the stability of the Product CS0980, Chitinase Assay Kit, substrate solution (s)?

    The substrate solution(s) should be stored on ice during the experiment.  For storage up to one month, freeze at -20°C or below.

  9. How many reactions will each substrate in Product CS0980, Chitinase Assay Kit, provide?

    If used at a concentration of 1mg/mL: *  4-Nitrophenyl N-acetyl-b-D-glucosaminide (Product No. N9376) -100 reactions; * 4-Nitrophenyl N,N’-diacetyl-b-D-chitobioside (Product No. N6133) -50 reactions; * 4-Nitrophenyl-b-D-N,N',N''-triacetylchitotriose (Product No. N8638) -10 reactions.

  10. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

Ahmed Akrem et al.
Acta crystallographica. Section F, Structural biology and crystallization communications, 67(Pt 3), 340-343 (2011-03-12)
A chitinase has been isolated and purified from Crocus vernus corms. N-terminal amino-acid sequence analysis of the approximately 30 kDa protein showed 33% identity to narbonin, a seed protein from Vicia narbonensis L. The C. vernus chitinase was crystallized by the
Functional characterization of Clostridium difficile spore coat proteins
Permpoonpattana P, et al.
Journal of Bacteriology, 195, 1492?1503-1492?1503 (2013)
E Patrick Fuerst et al.
Frontiers in plant science, 8, 2259-2259 (2018-02-08)
Seeds have well-established passive physical and chemical defense mechanisms that protect their food reserves from decay-inducing organisms and herbivores. However, there are few studies evaluating potential biochemical defenses of dormant seeds against pathogens. Caryopsis decay by the pathogenic Fusarium avenaceum
Ante Jerkovic et al.
Plant physiology, 152(3), 1459-1470 (2010-01-12)
Bran from bread wheat (Triticum aestivum 'Babbler') grain is composed of many outer layers of dead maternal tissues that overlie living aleurone cells. The dead cell layers function as a barrier resistant to degradation, whereas the aleurone layer is involved
Isolation, purification, crystallization and preliminary crystallographic studies of a chitinase from Crocus vernus
Akrem,A et al.
Acta Crystallographica Section A, Foundations of Crystallography, F67, 340-343 (2011)
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