"Xylanase" Related Products

Xylanase Assay Kit (Azo-Wax)

Cat.No.: Kit-0848
Product Overview: The Xylanase (Azo-Wax) test kit is suitable for the measurement and analysis of endo-1,4-β-D-xylanase in enzyme preparations, bread improver mixtures and animal feeds. Containing Azo-wheat arabinoxylan and a Trichoderma sp. xylanase control.
Size: 200 assays
Kit Components: 1. Azo-wheat arabinoxylan (100 mL, 1% w/v).
2. Trichoderma sp. control xylanase [3700 milli-Units (mU)/mL; pH 6.0 and 40°C] in 50% glycerol.
Materials Required but Not Supplied: EXTRACTION BUFFERS (not enclosed):
(A) Sodium acetate buffer (100 mM, pH 4.7)
Add 6.0 g of glacial acetic acid (1.05 g/mL) to 800 mL of distilled water. Adjust the pH to 4.7 with 2 M (8 g/100 mL) sodium hydroxide solution, and the volume to 1 litre. Add 0.2 g of sodium azide and dissolve. Stable at room temperature for > 12 months.
(B) MES buffer (100 mM) plus SDS (1% w/v)
Add 19.5 g of MES free acid (Sigma M-8250) to 800 mL of distilled water and dissolve. Adjust the pH to 6.0 with 1 M sodium hydroxide. Add 10 g of SDS (sodium lauryl sulphate; Sigma L-4509) and dissolve. Adjust the volume to 1 litre and add 0.2 g of sodium azide and dissolve. Stable at room temperature for > 12 months.
EQUIPMENT (Recommended):
1. Glass test tubes (round bottomed; 16 x 100mm and 16 x 120mm).
2. Micro-pipettor, e.g. Gilson Pipetman® (200 μL and 500 μL).
3. Positive displacement pipettor e.g. Eppendorf Multipette®- with 5.0 mL Combitip® (to dispense 0.5 mL aliquots of Azo-WAX solution).
- with 25 mL Combitip® (to dispense 2.5 mL aliquots of IMS or ethanol).
4. Analytical balance.
5. Spectrophotometer set at 590 nm.
6. Vortex mixer (e.g. IKA® Yellowline Test Tube Shaker TTS2).
7. Stop clock.
8. Whatman No.1 (9 cm) filter papers.
Detection method: Based on use of Azo-WAX reagent (590 nm)
Compatible Sample Types: Animal feeds, enzyme preparations, bread improver mixtures and other materials
Assay Protocol: EXTRACTION AND ASSAY OF XYLANASE IN FEED SAMPLES:
Trichoderma sp. Xylanases:
EXTRACTION:
1. Mill feed samples (approx. 50 grams) to pass a 0.5 mm screen and mix thoroughly.
2. Weigh 0.5 g ± (0.01 g) of each sample in duplicate into glass testtubes (16 x 120 mm).
3. Add 5 mL of 100 mM sodium acetate buffer (pH 4.7) to each sample and stir on a vortex mixer. Add 0.2 mL of water to one tube and 0.2 mL of xylanase (~ 740 mU) to the other. Immediately stir the slurries on a vortex mixer.
4. Incubate the slurries at room temperature and stir occasionally over the following 20 min.
5. Centrifuge the tubes at 1,500 g for 10 min in a bench centrifuge and use the supernatant directly in assays. Start assays within 30 min of obtaining these extracts to minimise loss of enzyme activity.
ASSAY:
1. Accurately transfer 0.5 mL aliquots of supernatant solutions (in duplicate) to glass test-tubes (16 x 100 mm) at room temperature.
2. Add 0.5 ml of Azo-WAX (1% w/v) to each tube and stir the tube vigorously. Immediately place the tubes in a water bath and incubate at 50°C ± 0.1°C for exactly 30 min.
3. Add 2.5 mL of IMS or ethanol and stir the tube vigorously on a vortex mixer. Store the tube at room temperature for 5 min. 3 This treatment terminates the reaction and precipitates non-depolymerised dyed substrate.
4. Centrifuge the tubes at 1,500 g for 10 min.
5. Measure the absorbance of the supernatant solutions at 590 nm against a reaction blank.
Prepare a Reaction Blank by adding 2.5 mL of IMS or ethanol to a mixture of 0.5 mL of Azo-WAX and 0.5 mL of 100 mM sodium acetate buffer (pH 4.7). Stir the slurry and store at room temperature for 5 min before centrifugation (1,500 g, 10 min). A single reaction blank is required for each feed sample.
Assay time: ~ 45 min
Analysis: The level of xylanase in the flour sample is calculated as follows:
Activity in feed sample (0.5 g) = Added activity x [SA/(TA - SA)]
where:
Added activity = the amount of xylanase added to the feed slurry at the time of assay e.g.: 740 mU in the control xylanase solution (0.2 mL).
SA = the reaction absorbance obtained for extracts of the feed to which no control xylanase was added.
TA = the total absorbance, i.e. the absorbance of extracts of the sample to which the control xylanase was added.
Sensitivity: 0.2 U/mL of assay solution
Products Types

◆ Assay kits

Online Inquiry

Note: There will be extra charge for optional service!

Please input "biomart" as verification code. Please review Creative BioMart's privacy policy for more information

Optional requirements on this protein    +Expand

Price Inquiry

Welcome! For price inquiries, please feel free to contact us through the form below. We will get back to you as soon as possible.