a-D-葡萄糖醛酸酶检测试剂盒 α-Glucuronidase Assay Kit 货号:K-AGLUA Megazyme试剂盒

a-D-葡萄糖醛酸酶检测试剂盒

英文名:α-Glucuronidase Assay Kit

货号:K-AGLUA

规格:50 Assays per kit

市场价: 3800

The Alpha-D-Glucuronidase test kit is a simple, reliable and accurate method for the measurement and analysis of alpha-D-glucuronidase in various enzyme preparations. Containing and aldouronic acid substrate and an α-D-glucuronidase control.
Suitable for manual and microplate formats.

UV-method for the measurement of α-D-Glucuronidase in
various enzyme preparations

Principle:
(α-D-glucuronidase)
(1) Aldouronic acid (tri:tetra:penta) + H2O →
β-(1,4)-D-xylo-oligosaccharides + D-glucuronic acid

(uronate dehydrogenase; UDH)
(2) D-Glucuronic acid + NAD+ + H2O → D-glucarate + NADH + H+

Kit size: 50 assays (manual) / 200 (microplate)
Method: Spectrophotometric at 340 nm
Reaction time: ~ 25 min
Detection limit: 17 mU/mL
Application examples:
Enzyme preparations and other materials
Method recognition: Novel method

Advantages

  • Very competitive price (cost per test)
  • All reagents stable for > 2 years as supplied
  • Simple format
  • Mega-Calc™ software tool is available from our website for hassle-free raw data processing
  • Standard included
  • Suitable for manual, microplate and auto-analyser formats

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木聚糖酶检测试剂盒 Xylanase Assay Kit (Azo-Wax) 货号:K-AZOWAX Megazyme试剂盒

木聚糖酶检测试剂盒

英文名:Xylanase Assay Kit (Azo-Wax)

货号:K-AZOWAX

规格:200 assays per kit

市场价: 5200

产品名称:木聚糖酶检测试剂盒
英文名称:Xylanase Assay Kit (Azo-Wax)

型号规格:200次
品牌:爱尔兰Megaeyme公司

自然界中,D-木糖以多糖形式如木聚糖,阿拉伯糖基木聚糖,GAX(glucuronoarabinoxylan),木葡聚糖和XGA(xylogalacturonan)存在。一些海藻中也存在交联D-木糖,据认定一些相近多糖构成洋车前子胶的骨架。番石榴,梨,黑莓,罗甘莓,悬钩子,芦荟维拉胶,褐藻,紫锥菊,乳香树,花椰菜,菠菜,茄子,豌豆,青豆,秋葵,卷心菜和谷物中存在游离的D-木糖。D-木糖用于小肠疾病诊断中的吸收实验,如食品中营养,维生素和矿物质吸收障碍。D-木糖可以被小肠正常轻松地吸收,当患有这些疾病时,D-木糖不能被吸收,血液和尿中的含量很低。D-木糖检测可以帮助确定儿童无法增重的原因,尤其是那些进食足够的儿童。如果在多糖中, D-木糖与其他糖类的比例已知,就可以通过酸水解测定D-木糖的浓度进而定量多糖。木聚糖是一些多糖的主要部分,这些多糖水解后的可发酵糖类可用于生产生物燃料。

原理

D-木糖的α-和β-变旋异构形式互换由木糖变旋酶(XMR)催化(1)。

在pH7.5下,β-D-木糖由β-木糖脱氢酶(β-XDH)催化,被NAD+氧化为D-木糖酸。

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.

Colourimetric method for the determination of Xylanase in feed,
foodstuffs and other materials

Principle:
(β-xylanase)
(1) Azo-WAX + H2O → Azo-WAX fragments
(insoluble in aqueous alcohol) (soluble in aqueous alcohol)

Kit size: 200 assays
Method: Based on use of Azo-WAX reagent (590 nm)
Total assay time: ~ 45 min
Detection limit: 0.2 U/mL of assay solution
Application examples:
Animal feeds, enzyme preparations, bread improver mixtures and
other materials
Method recognition:
Used widely in the feed industry

Advantages

  • Very cost effective
  • All reagents stable for > 2 years
  • Only test kit available
  • Simple format
  • Standard included

 

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内切纤维素酶检测试剂盒[CELLG3方法] Cellulase Assay Kit (CELLG3 Method) 货号:K-CELLG3 Megazyme试剂盒

内切纤维素酶检测试剂盒[CELLG3方法]

英文名:Cellulase Assay Kit (CELLG3 Method)

货号:K-CELLG3

规格:180 / 360 assays per kit / 720 (auto-analyser)

市场价: 6000

 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


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内切纤维素酶检测试剂盒[CELLG5方法] Cellulase Assay Kit (CELLG5 Method) 货号:K-CELLG5-2V Megazyme试剂盒

内切纤维素酶检测试剂盒[CELLG5方法]

英文名:Cellulase Assay Kit (CELLG5 Method)

货号:K-CELLG5-2V

规格:60 / 120 assays (manual) / 240 (auto-analyser)

市场价: 3800

The CELLG5 assay reagent for the measurement of
endo-cellulase (endo-1,4-β-glucanase) contains two components;
1) 4,6-O-(3-Ketobutylidene)-4-nitrophenyl-β-D-cellopentaoside (BPNPG5) and 2) thermostable β-glucosidase. The ketone blocking group prevents any hydrolytic action by the
β-glucosidase on BPNPG5. 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 4-nitrophenol is therefore directly related to the hydrolysis of BPNPG5 by the endo-cellulase. The reaction is terminated and the phenolate colour is developed on addition of Tris buffer solution (pH 9.0).

 

Colourimetric method for the determination of
endo
-1,4-β-glucanase (cellulase) in enzyme preparations and fermentation products

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

Advantages

  • Very cost effective
  • All reagents stable for > 4 years
  • Completely specific for cellulase (endo-1,4-glucanase)
  • Generally applicable and highly sensitive
  • Simple format. Well suited to automation
  • Standard included

 

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内切纤维素酶检测试剂盒[CELLG5方法] Cellulase Assay Kit (CELLG5 Method) 货号:K-CELLG5-4V Megazyme试剂盒

内切纤维素酶检测试剂盒[CELLG5方法]

英文名:Cellulase Assay Kit (CELLG5 Method)

货号:K-CELLG5-4V

规格:120 / 240 assays (manual) / 480 (auto-analyser)

市场价: 6042

The CELLG5 assay reagent for the measurement of
endo-cellulase (endo-1,4-β-glucanase) contains two components;
1) 4,6-O-(3-Ketobutylidene)-4-nitrophenyl-β-D-cellopentaoside (BPNPG5) and 2) thermostable β-glucosidase. The ketone blocking group prevents any hydrolytic action by the
β-glucosidase on BPNPG5. 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 4-nitrophenol is therefore directly related to the hydrolysis of BPNPG5 by the endo-cellulase. The reaction is terminated and the phenolate colour is developed on addition of Tris buffer solution (pH 9.0).

 

Colourimetric method for the determination of
endo
-1,4-β-glucanase (cellulase) in enzyme preparations and fermentation products

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

Advantages

  • Very cost effective
  • All reagents stable for > 4 years
  • Completely specific for cellulase (endo-1,4-glucanase)
  • Generally applicable and highly sensitive
  • Simple format. Well suited to automation
  • Standard included

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Alpha淀粉酶/液化酶/淀粉糖化酶检测试剂盒 Ceralpha: Alpha-Amylase Assay Kit 货号:K-CERA Megazyme试剂盒

Alpha淀粉酶/液化酶/淀粉糖化酶检测试剂盒

英文名:Ceralpha: Alpha-Amylase Assay Kit

货号:K-CERA

规格:100 assays per kit

市场价: 4452

AOAC Method 2002.01, AACC Method 22.02.01, ICC Standard No. 303, RACI Standard Method, CCFRA Flour Testing Working Group Method 0018. For the specific measurement of α-amylase in cereal grains and fermentation broths (fungal and bacterial).

Alpha淀粉酶/液化酶/淀粉糖化酶检测试剂盒

Q1. We would like to obtain an assay capable of quantifying trace amounts of alpha-amylase activity in the pullulanase preparation. Please let me know if Megazyme has any suitable assay available.

We think the Ceralpha Reagent would probably be best for this application.

Q2. I am interested in the Amylazyme and Ceralpha available from Megazyme. Are they applicable not only for cereal and microbial alpha-amylase but also porcine pancreatic alpha-amylase?

The Ceralpha method is excellent for all alpha-amylases and will work fine for porcine pancreas alpha-amylase at pH 6.9.

Q3. We would like to know where the coefficient of 18.1 (EmM of p-nitrophenol) comes from? What does it mean?

This is the extinction coefficient, i.e. absorbance of a 1 mM solution of p-nitrophenol in a 1 cm light path at 400 nm.

Q4. I wish to measure the enzyme activity of alpha-amylase in bread after cooking. Will your Ceralpha Amylase Kit work on cooked breads? If so, do I need to alter the procedure?

The level of alpha-amylase in wheat flour is quite low, so in bread baked from this flour it is likely to be much lower.  The Ceralpha test is very sensitive, but you can increase sensitivity by increasing incubation time up to several hours (6 hours).  This should allow measurement of the enzyme if there is any there.

Q5. Any recommendations on using your Ceralpha alpha-amylase test at a low pH (pH 3-4)?

Cereal alpha-amylase assay reagent cannot be used at pH 3-4.  However, you can perform a similar assay using blocked p-nitophenol maltoheptaoside (BPNPG7). Basically dissolve the BPNPG7 vial contents in 10 mL with water.  Incubate 0.2 mL of your enzyme (buffered) with 0.2 mL of BPNPG7 for 10 min at a set temperature. Terminate the reaction by heating in a boiling water bath for 2 min.  Then add 0.2 mL of alpha-glucosidase (E-TSAGL; at 10 U/mL in 0.2 M tris buffer, pH 7.0) and incubate for 10 min.  Add 3 mL stopping reagent (Trizma base pH 8.5) and read colour at 400 nm.

Q6. Can the Ceralpha kit distinguish between different forms of alpha-amylase?

The Ceralpha Kit does not distinguish between the different forms of alpha amylase, however some idea of relative levels of fungal and bacterial alpha-amylase can be obtained by performing the assay at different pH values.  Please refer to the Ceralpha kit booklet on this web site.

Q7. In order to use the Ceralpha kit to test for the presence of alpha-amylase, does one have to mill the wheat and if so can I grind the grain in a blender?

Cereal grains do need to be milled to allow effective extraction of alpha-amylase (and other constituents).  We recommend milling to pass a 0.5 mm screen.  You may have some success with a cheap coffee grinder.  However, the reproducibility will not be as good (it may be adequate for your requirements).

Q8. Can the Ceralpha reagent be used at high temperatures?

The Ceralpha method is routinely run at 40˚C, but it can be used at temperatures up to 60˚C.  If it is run at higher temperatures (up to 60˚C) then higher activity values will be obtained.  This assumes that the enzymes being analysed (alpha-amylase) are stable at these higher temperatures.  If you need to measure activity at temperatures above 60˚C you can use the Megazyme Amylazyme tablets or Red Starch.

Q9. Can Ceralpha Method determine the alpha-amylase activity in a protease preparation?

The standard method should be fine.  You can best check this by performing a time course hydrolysis of the alpha-amylase.  This should be linear if the alpha-glucosidase in the kit reagent is not attacked by the protease.

Q10. Can the internal glycosidic linkages only be cleaved by alpha-amylase?

Yes, only alpha-amylase can cleave the internal glycosidic bonds.

Q11. After alpha-amylase cleaves the internal glycosidic linkage, can beta-amylase or glucoamylase accelerate the hydrolysis reaction of the released p-nitrophenyl maltosaccharide?

When alpha-amylase cleaves the glycosidic linkage in the blocked substrate, any other enzymes in the extract do not accelerate the hydrolysis to give free p-nitrophenol.  The reason for this is that the level of thermostable alpha-glucosidase in the substrate mixture is saturating.

Q12. Can Rice alpha-amylase be analysed by Ceralpha kit?

Rice alpha-amylase can be assayed with Ceralpha kit.  Alpha-glucosidase in the rice extract will not interfere.

Q13. Why is extraction conducted at room temperature or 40˚C, and not at 4˚C?

We have found that we get effective extraction from a range of cereal flours at room temperature (approx. 22˚C) and that the enzyme was very stable at this temperature for several hours.  For wheat flour, the optimal temperature for extraction is 40˚C (refer to the Ceralpha Booklet).

Q14. Can the Ceralpha method be used to determine alpha-amylase from oat flour extracts?

The Ceralpha method can be used for oats.  The only likely problem is the high viscosity of the extract.  If the extract looks very viscous, double the ratio of extraction buffer to flour.  Allow for this in the calculations.  If the absorbances are low then increase the incubation time to say 30 minutes.  Then allow for this increased time of incubation in the calculations.

Q15. Is there a method to increase the sensitivity of assay regarding the measurement of alpha-amylase in fermentation broth?

We suggest that the fermentation broth is simply adjusted to 5.0 with acid or base and this can be used directly in the assay. The sensitivity can then be increased by increasing incubation time up to 4 hours.  Appropriate adjustments then need to be made to the calculations.

Q16. Can the reaction blank be used to zero the spectrophotometer directly?

You can zero the spectrophotometer directly with the blank.  However, it is wise to know how high the blank is to ensure that the substrate is OK, i.e. the blank is usually 0.05-0.06.  If it is above 0.1, it has probably been contaminated with some alpha-amylase enzyme.

Q17. When the absorbance is between 1.0–1.5, can the solution be diluted directly with an additional 3.0 mL of distilled water to cut the absorbance in half before reading it?

If the absorbance value is above 1.2 then the assay will be limited by the amount of available substrate.  Thus, you cannot simply dilute the colour.

Q18. I have immobilised alpha-amylase onto glass bead and now I want to test the activity, can the Ceralpha kit test for this?

To measure alpha-amylase on glass beads I would recommend the use of the Ceralpha method.  The kit is complete and extremely easy to use.  Perhaps to assay, you may freeze dry a small sample of the beads and then weigh an appropriate amount (say 5 mg) into a test tube.  Add 0.2 mL of buffer pH 5-6 and 0.2 mL Ceralpha reagent.  Incubate at 40˚C for 10 min.

Q19. Is it possible to measure acid-stable alpha-amylase by the Ceralpha Kit?

The Ceralpha method should be fine for acid stable alpha-amylase.  However, assays should be performed in the pH range of 5.2 to 7.5.

Q20. Can you tell me the application of the Amylazyme Tabs and Ceralpha Kit to residual enzymes in finished bakery products, particularly in sweet goods?

For measurement of residual enzymes in bakery products, you should use the Amylazyme method rather than the Ceralpha method.  With the standard assay formats available, the sensitivity of the Amylazyme method is 10-20 times greater than can be achieved with the Ceralpha method.  You will need this greater sensitivity to measure the extremely low levels of activity present.

Q21. What would be an expected level of Ceralpha units in germinated or malted barley?

The level of alpha-amylase in malted barley is about 150 Ceralpha units per gram, which would be the expected level in germinated barley grain.

Q22. Should the pH of the sample be adjusted even for samples in acidic media?

The pH of the assay solution after the sample is added should be the same as that of the assay buffer that is supplied with the kit.
Low sample volumes (e.g. 0.1 mL) are not likely to affect the pH of the assay solution and therefore may not require pH adjustment.

Megazyme 溶解淀粉 操作视频

Megazyme 试剂盒样品前处理准备操作视频

Megazyme α淀粉酶检测试剂盒Ceralpha操作视频(K-CERA)

葡萄糖氧化酶检测试剂盒 Glucose Oxidase Assay Kit 货号:K-GLOX Megazyme试剂盒

葡萄糖氧化酶检测试剂盒

英文名:Glucose Oxidase Assay Kit

货号:K-GLOX

规格:200 assays (manual) / 2000 assays (microplate)

市场价: 2862

The Glucose Oxidase assay kit is a simple procedure for the rapid and reliable measurement and analysis of glucose oxidase activity in industrial enzyme preparations and bread improver mixtures.

Glucose Oxidase assay kit is suitable for manual, auto-analyser and microplate formats.
 

Colourimetric method for the determination of Glucose Oxidase
in foodstuffs and fermentation products

Principle:
(glucose oxidase)
(1) D-Glucose + H2O + O2 → D-glucono-δ-lactone + H2O2

(peroxidase)
(2) 2H2O2 + p-hydroxybenzoic acid + 4-aminoantipyrine →
quinoneimine + 4H2O

Kit size: 200 assays (manual) / 2000 (microplate)
/ 1960 (auto-analyser)
Method: Spectrophotometric at 510 nm
Reaction time: ~ 20 min
Detection limit: 10 U/L
Application examples:
Enzyme preparations, and other materials (e.g. biological cultures,
samples, etc.)
Method recognition: Novel method

Advantages

  • Very competitive price (cost per test)
  • All reagents stable for > 12 months after preparation
  • Simple format
  • Mega-Calc™ software tool is available from our website for hassle-free raw data processing
  • Standard included
  • Suitable for manual, microplate and auto-analyser formats

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麦芽淀粉酶检测试剂盒 Malt Amylase Assay Kit 货号:K-MALTA Megazyme试剂盒

麦芽淀粉酶检测试剂盒

英文名:Malt Amylase Assay Kit

货号:K-MALTA

规格:100 assays (50 of each) per kit

市场价: 4452

The Malt Amylase test kit is suitable for the specific measurement and analysis of α-amylase and of β-amylase in malt flour.

Colourimetric method for the determination of α-Amylase and
β-Amylase in cereal grains, malt, food, beverages and
fermentation products

Principle:
(1) α-Amylase is measured using the “Ceralpha” Method as used
in K-CERA

(2) β-Amylase is measured using the “Betamyl-3” Method as used
in K-BETA3

Kit size: 50 assays of each
Method: Spectrophotometric at 400 nm
Reaction time: ~ 20 min (Ceralpha Method)
~ 10 min (Betamyl-3 Method)
Detection limit: 0.05 U/mL
Application examples:
Cereal flours, malts, fermentation broths and other materials
Method recognition:
“Ceralpha” Method: AOAC (Method 2002.01), AACC (Method
22-02.01), ICC (Standard No. 303), RACI
(Standard Method) and CCFRA (Flour Testing
Working Group Method 0018)
“Betamyl-3” Method: RACI (Standard Method)

Advantages

  • Very cost effective
  • All reagents stable for > 2 years as supplied
  • Only enzymatic kit available (Beta-Amylase)
  • Very specific
  • Simple format
  • Mega-Calc™ software tool is available from our website for hassle-free raw data processing
  • Standard included

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Beta葡聚糖酶[麦芽和微生物]检测试剂盒 β-Glucanase Assay Kit (Malt and Microbial) 货号:K-MBGL Megazyme试剂盒

Beta葡聚糖酶[麦芽和微生物]检测试剂盒

英文名:β-Glucanase Assay Kit (Malt and Microbial)

货号:K-MBGL

规格:100 assays per kit

市场价: 4134

β-Glucanase (Malt and Microbial) Assay Kit is suitable for the measurement and analysis of malt and bacterial β-glucanase and endo-1,4-β-glucanase.

Colourimetric method for the determination of β-Glucanase
in malt, foodstuffs and fermentation products

Principle:
(β-glucanase)
(1) Azo-Barley β-glucan (polymer) → Azo-barley β-glucan
(fragments)

(2) Add alcohol; centrifuge to remove polymeric Azo-barley
β-glucan

(3) Measure the absorbance of the supernatant solution

Kit size: 100 assays
Method: Spectrophotometric at 590 nm
Reaction time: ~ 30 min
Detection limit: 100 U/kg of malt
Application examples:
Malt extracts, wort, beer and other materials
Method recognition:
RACI (Standard Method)

Advantages

  • Very cost effective
  • All reagents stable for > 2 years during use
  • Only kit available
  • Very specific
  • Simple format
  • Standard included

Q1. We have measured the molecular weight of beta-glucan originating from barley ordered from your company. Can you please tell us which method you have used for measuring molecular weight?

The MW’s were determined by Multiangle laser light scattering technique.

Q2. I have purchased barley beta-glucan (lot 30108) and carob galactomannan low viscosity (lot 30702) and would like to know what else there might be in these substrates.

Barley beta-glucan lot 30108 would contain about 3-4% arabinoxylan (this was produced in 1993).  Material supplied post 1995 contains < 0.5.% arabinoxylan. Carob galactomannan is quite pure (> 96%).

Q3. Although not specified on the beta-glucan data sheet, is the ratio of 1-3 to 1-4 bonds measured? If so, what is the ratio and would you expect it to remain standardised over a number of different batches?

The content of 1,3 bonds in barley beta-glucan is about 32% (from literature).  We would not expect this to change much (if at all) over different batches.

Q4. Both substrates, barley beta-glucan and wheat arabinoxylan, are standardised to a specific viscosity, e.g. 23-24 cSt. Are the substrates adjusted to give this viscosity?

The substrates are enzymically treated to yield the required viscosity (20 ~ 30 cSt). Modification of the viscosity of beta-glucan is required for IOB viscometric malt beta-glucanase test (which works well).

Q5. Should the pH of the sample be adjusted even for samples in acidic media?

The pH of the assay solution after the sample is added should be the same as that of the assay buffer that is supplied with the kit.
Low sample volumes (e.g. 0.1 mL) are not likely to affect the pH of the assay solution and therefore may not require pH adjustment.
Samples above 0.1 mL are more likely to affect the pH of the assay solution and therefore the pH of these samples should be adjusted as described in the data booklet, prior to addition to the assay.

Q6. There is an issue with the performance of the kit; the results are not as expected.

If you suspect that the Megazyme test kit is not performing as expected such that expected results are not obtained please do the following:

  1. Ensure that you have tested the standard sample that is supplied with the Megazyme test kit.
  2. Send the results of the kit standard, blank samples and the results obtained for your sample, in the relevant MegaCalc spreadsheet (if available) to Megazyme (cs@megazyme.com). Where available the relevant MegaCalc spreadsheet can be downloaded from where the product appears on the Megazyme website.
  3. State the kit lot number being used (this is found on the outside of the kit box).
  4. State which assay format was used (refer to the relevant page in the kit booklet if necessary).
  5. State exact details of any modifications to the standard procedure that is provided by Megazyme.
  6. State the sample type and describe the sample preparation steps if applicable.

Q7. We want to set up reducing-sugar assays for beta-glucanase and xylanase from different fungal origins. There are three kinds in your catalogue – high viscosity, medium viscosity and low viscosity. Which do you recommend for the glucanase assay.

For glucanase assay we recommend the medium viscosity beta-glucan.  We now offer low, medium and high viscosity wheat arabinoxylans, and we think that the low viscosity material will be best (easiest) to use in the reducing-sugar assay. 

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普鲁兰酶/极限醍醐精检测试剂盒 Pullulanase/Limit-Dextrinase Assay Kit (PULLG6 Method) 货号:K-PULLG6 Megazyme试剂盒

普鲁兰酶/极限醍醐精检测试剂盒

英文名:Pullulanase/Limit-Dextrinase Assay Kit (PULLG6 Method)

货号:K-PULLG6

规格:100 assays (manual)

市场价: 5194

PULLG6 assay for the measurement of pullulanase employs a water soluble defined substrate, namely 4,6-O-benzylidene-4-nitrophenyl-63-α-D-maltotriosyl-maltotriose (BPNPG3G3), coupled with the ancillary enzymes α-glucosidase and β-glucosidase. Upon hydrolysis of the substrate at the 1,6-α-linkage by pullulanase or limit-dextrinase, the released 4-nitrophenyl-β-maltotrioside is immediately hydrolysed to glucose and 4-nitrophenol by the concerted action of the α-glucosidase and β-glucosidase enzymes in the reagent mixture. The reaction is terminated and phenolate ions are developed by addition of dilute alkali. The absorbance is read at 400 nm and the value obtained correlates directly with pullulanase activity.

Colourimetric method for the determination of pullanase
or limit-dextrinase

Principle:
(pullulanase/limit-dextrinase)
(1) Benzylidene-G3-(α-1,6)-G3-β-PNP + H2O → Benzylidene-G3
+ G3-β-PNP

(thermostable α-glucosidase and β-glucosidase)
(2) G3-β-PNP + H2O → D-glucose + PNP

(alkaline solution)
(3) PNP → phenolate ion (yellow colour)

Note: PNP = 4-nitrophenol

Kit size: 100 assays
Method: Spectrophotometric at 400 nm
Total assay time: 10 min for pullanase preparations
30 min for malt extracts containing
limit-dextrinase
Detection limit: 0.18 U/mL for pullulanase preparations
(50-fold dilution)
0.01 U/g for limit dextrinase in milled malt
Application examples: Assay of microbial pullulanase preparations
Measurement of limit-dextrinase in
malt extracts
Method recognition: Novel method

Advantages

  • High sensitivity
  • Suitable for manual and auto-analyser formats
  • No transglycosylation interference
  • Very cost effective
  • All reagents stable for > 1 year after preparation
  • Very specific

  • Simple format

  • Standard included

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α淀粉酶SD检测试剂盒(高感光度法) α-Amylase SD Assay Kit (High Sensitivity Method) 货号:K-AMYLSD Megazyme试剂盒

α淀粉酶SD检测试剂盒(高感光度法)

英文名:α-Amylase SD Assay Kit (High Sensitivity Method)

货号:K-AMYLSD

规格:160 / 320 assays (manual) / 640 assays (auto-analyser)

市场价: 4346

The Amylase SD Method: A highly sensitive colourimetric method for the determination of α-amylase in sprout damaged wheat grain (also known as pre-harvest sprouting or weather damaged wheat grain) and “late maturity α-amylase” wheat grain.

Advantages

  • Extremely high sensitivity – 2.4-fold increase over Ceralpha (K-CERA)
  • Very cost effective
  • All reagents stable for > 2 years after preparation
  • Very specific
  • Simple format
  • Mega-Calc™ software tool is available from our website for hassle-free raw data processing
  • Standard included

  • Suitable for Maual and auto-analyser formats

Highly sensitive colourimetric method for the determination of α-Amylase in sprout damaged grain

Principle:
(α-amylase)
(1) Ethylidene-G7-α-PNP + H2O → Ethylidene-GX + G(7-X)-α-PNP

(thermostable α-glucosidase)
(2) G(7-X)-α-PNP + H2O → D-glucose + PNP

(alkaline solution)
(3) PNP → phenolate ion (yellow colour)
Note: PNP = 4-nitrophenol

Kit size: 160 / 320 assays (manual) / 640 (auto-analyser)
Method: Spectrophotometric at 400 nm
Total assay time: ~ 5 min
Detection limit: 0.05 U/mL
Application examples: Sprout damaged wheat grain
Method recognition: Novel method

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阿拉伯树胶二糖[纯度>95%][浆] Arabinobiose (syrup) – 50mg 货号:O-ABI Megazyme试剂盒

阿拉伯树胶二糖[纯度>95%][浆]

英文名:Arabinobiose (syrup) – 50mg

货号:O-ABI

规格:50 mg

市场价: 2968

CAS: 78088-21-8
Molecular Formula: C10H18O9
Molecular Weight: 282.2
Purity: > 95%

High purity Arabinobiose (syrup) for use in research, biochemical enzyme assays and in vitro diagnostic analysis.

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2-氯-4-硝基苯基β-纤维四糖 2-Chloro-4-nitrophenyl-β-cellotetraoside 货号:O-CPNPG4-50 Megazyme试剂盒

2-氯-4-硝基苯基β-纤维四糖

英文名:2-Chloro-4-nitrophenyl-β-cellotetraoside

货号:O-CPNPG4-50

规格:50 mg

市场价: 6000

CAS: 189094-93-7
Molecular Formula: C30H44CINO23
Molecular Weight: 822.1
Purity: > 93%

High purity 2-Chloro-4-nitrophenyl-β-cellotetraoside for use in research, biochemical enzyme assays and 
in vitro diagnostic analysis.

A potential substrate for the measurement of cellulase (endo-1,4-β-glucanase).       

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阿拉伯树胶七糖[纯度>95%][粉] Arabinoheptaose (powder) – 15mg 货号:O-AHP Megazyme试剂盒

阿拉伯树胶七糖[纯度>95%][粉]

英文名:Arabinoheptaose (powder) – 15mg

货号:O-AHP

规格:15 mg

市场价: 2300

CAS: 190852-27-8
Molecular Formula: C35H58O29
Molecular Weight: 924.8
Purity: > 95%

High purity Arabinoheptaose (powder) for use in research, biochemical enzyme assays and in vitro diagnostic analysis.

Purity ~ 95%

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阿拉伯树胶六糖[纯度>95%][粉] Arabinohexaose (powder) – 20mg 货号:O-AHE Megazyme试剂盒

阿拉伯树胶六糖[纯度>95%][粉]

英文名:Arabinohexaose (powder) – 20mg

货号:O-AHE

规格:20 mg

市场价: 2438

CAS: 190852-26-7 
Molecular Formula: C30H50O25
Molecular Weight: 810.7
Purity: > 95%



High purity Arabinohexaose (powder) for use in research, biochemical enzyme assays and in vitro diagnostic analysis.

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