纤维素酶是一种重要的酶产品，是一种复合酶，主要由外切β-葡聚糖酶、内切β-葡聚糖酶和β-葡萄糖苷酶等组成，还有很高活力的木聚糖酶。
提供内切纤维素酶检测试剂盒，色度法（400 nm）测定酶制品和发酵产品中的纤维素酶（内切-1,4-β-葡聚糖酶）。
内切纤维素酶检测试剂盒(CELLG3方法) K-CELLG3
使用高纯度β-葡萄糖苷酶和苯亚甲基阻断，2 – 氯-4 – 硝基苯基-β-Dcellotrioside（BClPNPβ-G3）。 β-葡萄糖苷酶的能力确保测定的可靠的最大的灵敏度。BClPNPβ-G3的水解为苯亚甲基，通过纤维素酶阻断纤维二糖和2-氯-4 -硝基苯基-β-D-葡萄糖， 2-Cl-4-硝基苯基-β-D-葡萄糖通过β-葡萄糖苷酶立即裂解为D-葡萄糖和游离的2 – 氯-4 – 硝基苯酚（ClPNP）。
反应时间：16min
适用样品：酶制品和发酵产品
优点：
价格低廉（每次检测成本）
特异性高
方法简单
包含标准品
The CELLG3 assay reagent for the measurement of endo-cellulase (endo-1,4-β-glucanase) contains two components;
1) 4,6-O-benzylidene-2-chloro-4-nitrophenyl-β-D-cellotrioside (BCNPG3) and 2) thermostable β-glucosidase. The benzylidene blocking group prevents any hydrolytic action by the β-glucosidase on BCNPG3. Incubation with an endo-cellulase generates a non-blocked colourimetric oligosaccharide that is rapidly hydrolysed by the ancillary β-glucosidase. The rate of formation of 2-chloro-4-nitrophenol is therefore directly related to the hydrolysis of BCNPG3 by the endo-cellulase. The reaction is terminated and the phenolate colour is developed on addition of Tris buffer solution (pH 9.0).
Please note that a new assay kit (K-CELLG5) is now available for the measurement of endo-cellulase. The CELLG5 reagent contains a cellopentaose core and exhibits vastly improved sensitivity for some cellulases. In addition, the exchange of the benzylidene blocking group in CELLG3 for 3-keto-butylidene in CELLG5 improves the substrate’s water solubility significantly, allowing for a reduction in the concentration of DMSO required in the assay. As DMSO is known to inhibit certain cellulases, this is another benefit in using CELLG5. Megazyme now recommends the use of K-CELLG5 for all assays for the measurement of endo-cellulase.
Colourimetric method for the determination of
endo-1,4-β-glucanase (cellulase) in enzyme preparations and fermentationproducts
Principle:
(
endo-1,4-β-glucanase)
(1) 3-Ketobutylidene-G5-β-PNP + H2O → Blocked-GX + G(5-X)-β-PNP
(thermostable β-glucosidase)
(2) G(5-X)-β-PNP + H2O → D-glucose + PNP
(alkaline solution)
(3) PNP → phenolate ion (yellow colour)
Note:
PNP = 4-nitrophenol
Kit size:
K-CELLG5-4V 120 / 240 assays (manual) / 480 (auto-analyser)
or
K-CELLG5-2V 60 / 120 assays (manual) / 240 (auto-analyser)
Method: Spectrophotometric at 400 nm
Total assay time: 10 min
Detection limit: 3.5 x 10-4 U/mL
Application examples:
Fermentation broths, industrial enzyme preparations, biofuels research
Method recognition: Novel method
睡眠研究用
抗小鼠Trβ2，兔
抗小鼠皮卡丘素，兔
