Chitinase Assay Kit, Fluorimetric
SIGMA/CS1030 - sufficient for 200 multiwell tests
Synonym: Chitinase Detection Kit
Product Type: Chemical
Photo is meant to be representative of packaging you will receive if ordered. Appropriate product information is printed on actual labels and package types are subject to change.
Unit Definition One unit will liberate 1.0 mg of N-acetyl-D-glucosamine from chitin per hour at pH 6.0 at 25 °C.
Chitinases have been detected in many organisms, including bacteria, fungi, plants, invertebrates and vertebrates. Chitinases are broadly classified as endo- and exochitinases. The endochitinase activity is defined as the random cleavage at internal points in the chitin chain. The exochitinase activity is defined as the progressive action starting at the non reducing end of chitin with the release of chitobiose or N-acetylglucosamine units. Chitobiosidase and N-acetyl-b-glucosaminidase are considered exochitinases. The combination of endo- and exochitinases results in a synergistic increase in the chitinolytic activity.
| Quality Level | 200  |
| shipped in | wet ice |
| storage temp. | −20°C |
| usage | sufficient for 200 multiwell tests |
| 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-chitobi • 4-Methylumbelliferyl N-acetyl-β-D-glucosaminid • 4-Methylumbelliferyl β-D-N,N′,N′′-triacetylchi |
| Application: | 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. |
| 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 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 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. |
| Preparation Note: | Use ultrapure water for preparation of reagents. |
| Symbol |  GHS07 |
| Signal word | Warning |
| Hazard statements | H319 |
| Precautionary statements | P305 + P351 + P338 |
| Hazard Codes | Xi |
| Risk Statements | 36 |
| Safety Statements | 26 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 12161503 |
| Components | 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 |