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Merck

CS0980

Sigma-Aldrich

Chitinase Assay Kit

sufficient for 100 multiwell tests

Sinónimos:

Chitinase Activity Detection Kit

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About This Item

UNSPSC Code:
12161503
NACRES:
NA.54
En este momento no podemos mostrarle ni los precios ni la disponibilidad

usage

sufficient for 100 multiwell tests

Quality Level

absorbance ratio

405 nm (Absorbance)

shipped in

wet ice

storage temp.

2-8°C

Gene Information

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.

Application

Chitinase Assay Kit has been used in screening chitinase activity of recombinant chitinase from yeast, Metschnikowia fructicola,[1] bacterial endophyte 3A12[2] and Clostridium difficile spores.[3]

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.

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.

Solo componentes del kit

Referencia del producto
Descripción

  • Assay Buffer 20 mL

  • 4-Nitrophenyl N-acetyl-β-D-glucosaminide 10 mg

  • 4-Nitrophenyl N,N′-diacetyl-β-D-chitobioside 5 mg

  • 4-Nitrophenyl β-D-N,N′N′′-triacetylchitotriose 1 mg

  • Chitinase from Trichoderma viride 1 mg

  • p-Nitrophenol Solution, 10 mM 1 mL

  • Sodium Carbonate 1 g

related product

Referencia del producto
Descripción
Precios

pictograms

Health hazard

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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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
Genes required for the anti-fungal activity of a bacterial endophyte isolated from a corn landrace grown continuously by subsistence farmers since 1000 BC
Shehata HR, et al.
Frontiers in Microbiology, 7, 1548-1548 (2016)
Patima Permpoonpattana et al.
Journal of bacteriology, 193(23), 6461-6470 (2011-09-29)
Clostridium difficile is an important human pathogen and one where the primary cause of disease is due to the transmission of spores. We have investigated the proteins found in the outer coat layers of C. difficile spores of pathogenic strain
Postharvest application of a novel chitinase cloned from Metschnikowia fructicola and overexpressed in Pichia pastoris to control brown rot of peaches
Banani H, et al.
International Journal of Food Microbiology, 199, 54-61 (2015)
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)

Questions

1–8 of 8 Questions  
  1. Hello, I purchasee chitinase assay kit CS0980 last month. Can I use the kiy to analyse chitinase in hepatopancreas of lobsters? I dont find the procedure in the kit to prepare and measure chitinase from tissue samples

    1 answer
    1. As noted in its Technical Bulletin, the kit has been tested on Trichoderma viride, Streptomyces griseus and human macrophages. No protocol is available for the use of the kit to determine Chitinase activity in hepatopancreas of lobsters.

      Helpful?

  2. We are looking for established CHIT1enzyme activity assays for analysis of clinical samples (human serum or plasma).Does this kit apply to that?

    1 answer
    1. Yes, this kit is suitable for analyzing human serum and plasma. Please note that this product is for R&D use only, not for drug, household, or other uses.

      Helpful?

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

    1 answer
    1. 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.

      Helpful?

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

    1 answer
    1. 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).

      Helpful?

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

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

      Helpful?

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

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

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

    1 answer
    1. 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.

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

    1 answer
    1. 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.

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