淀粉葡萄糖苷酶试剂混合物 200次/瓶 4瓶爱尔兰

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产品名称：淀粉葡萄糖苷酶试剂混合物 200次/瓶 4瓶爱尔兰
产品型号：JKY/R-AMGR3
所属类别：
发布时间：2014-10-11
已获点击：366

详细说明

淀粉葡萄糖苷酶试剂混合物 200次/瓶 4瓶爱尔兰 型号:JKY/R-AMGR3

库号：M245787

对硝基苯酚,?-D-麦芽糖,?-葡萄糖苷酶

参数:
pH值 7.0

气味无

形态粉

稳定性冰箱内存十年或以上

成分:
名称 CAS比例
对硝基苯酚β-D-麦芽糖 95％

β-葡萄糖苷酶（杏仁） 5％

共4瓶,200次/瓶.

Amyloglucosidase Assay Reagent – 4 vials
p-Nitrophenyl ?-D-maltoside plus excess ?-glucosidase.

Catalogue Number: R-AMGR3
Content: 200 assays per vial

Appearance Off white powder
Specific Gravity Not applicable
Solubility in Water Readily soluble
pH Value 7.0
Odour none
Form powder
Stability stable in a freezer for ten or more years
Ingredients Name CAS Proportion
p-Nitrophenyl b-D-maltose 95%
b-Glucosidase (Almond) 5%

NOTE:
The AMG assay reagent previously supplied by Megazyme
contained p-nitrophenyl-β-maltoside (4 mM), plus almond seed
β-glucosidase (25 U/ml). While this substrate worked well, its
use was limited to a narrow pH and temperature range. Also,
stability was limited by the purity of the almond β-glucosidase
then available. Recently, this substrate has been improved by
replacing almond β-glucosidase with a thermostable
β-glucosidase. The advantages are:
1. The high purity of the thermostable β-glucosidase means that
the substrate is stable for longer periods of time.
2. Because the enzyme is thermostable, the reagent can be used
at temperatures up to 60°C (the preferred temperature for the
assay of AMG).
However, because thermostable β-glucosidase is unstable at
very low salt concentrations, salt and buffer are added to the
dry reagent. When dissolved, the substrate solution will have a
pH of ~ 6.0 (the optimal pH for stability of the β-glucosidase).

SUBSTRATE:
p-Nitrophenyl-β-maltoside (4 mM),
plus thermostable β-glucosidase (5 U/ml).
Dissolve the contents of one vial in 10 mL of distilled water, divide
into aliquots of 2-3 mL and store frozen. Store on ice during use.
BUFFER:
200 mM Sodium acetate buffer (pH 4.5).
Add 5.9 mL of glacial acetic acid (1.05 g/mL) to 900 mL of distilled
water. Adjust the pH to pH 4.4 by addition of 1 M (4 g/100 mL)
NaOH solution (approx. 30 mL is required). Adjust the volume to
1 L and store in a well sealed bottle at 4°C.
SAMPLE PREPARATION:
Add 1 mL of enzyme preparation to 9 mL of 200 mM sodium
acetate buffer (pH 4.5) and mix well. Repeat this dilution step
until the enzyme is suitably diluted for assay.
ASSAY PROCEDURE:
1. Pre-equilibrate enzyme solution at 40°C for 5 min.
2. To 0.2 ml of pre-equlibrated substrate solution add 0.2 mL
of suitably diluted (and pre-equilibrated) enzyme solution.
Mix well and incubate at 40°C for exactly 10 min.
1
NOTE:
The AMG assay reagent previously supplied by Megazyme
contained p-nitrophenyl-β-maltoside (4 mM), plus almond seed
β-glucosidase (25 U/ml). While this substrate worked well, its
use was limited to a narrow pH and temperature range. Also,
stability was limited by the purity of the almond β-glucosidase
then available. Recently, this substrate has been improved by
replacing almond β-glucosidase with a thermostable
β-glucosidase. The advantages are:
1. The high purity of the thermostable β-glucosidase means that
the substrate is stable for longer periods of time.
2. Because the enzyme is thermostable, the reagent can be used
at temperatures up to 60°C (the preferred temperature for the
assay of AMG).
However, because thermostable β-glucosidase is unstable at
very low salt concentrations, salt and buffer are added to the
dry reagent. When dissolved, the substrate solution will have a
pH of ~ 6.0 (the optimal pH for stability of the β-glucosidase).
3. Terminate the reaction and develop the colour by adding
3.0 mL of 2% trizma base solution (pH ~ 8.5; Sigma T-1503).
4. Measure the absorbance at 400 nm against a reagent blank.
CALCULATION OF ACTIVITY:
Activity (U/mL) = ΔA400 x 3.4 x 1 x Dilution.
10 0.2 18.1
where:
U = International units of enzyme activity. One Unit is the
amount of enzyme which release one μmole of
p-nitrophenol from the substrate per minute at the
defined pH and temperature.
ΔA400 = absorbance (reaction) - Absorbance (blank)
10 = incubation time
3.4 = final reaction volume (mL)
0.2 = volume of enzyme assayed (mL)
18.1 = EmM p-nitrophenol in 2% trizma base (pH ~ 8.5)
at 400 nm.
The Units of amyloglucosidase activity obtained using the
above assay, can be related to activity on maltose (10 mg/mL) or
soluble starch (10 mg/mL) at 40°C and pH 4.5, using the following
equations:
Enzyme Units on Maltose = 2.0 x Units on pNP-β-maltoside.
Enzyme Units on Starch = 15 x Units on pNP-β-maltoside.
REFERENCE:
McCleary, B.V., Bouhet, F. and Driguez, H. “Measurement
of amyloglucosidase using p-nitrophenyl β-maltoside as substrate”
(1991) Biotechnology Techniques, 5, 255-258.
2
NOTE:
The reagent blank is prepared by adding 3.0 mL of Trizma base
solution (2%) to 0.2 mL of reagent mixture with vigorous
stirring, followed by the enzyme solution (0.2 mL) with stirring

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Beta-Amylase Assay Reagent 100120002 Beta淀粉酶检测试剂 6 vials R-BAMR6

Ceralpha:alpha-Amylase Reagent 100120003 Alpha淀粉酶检测试剂 4 vials R-CAAR4

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东方化玻(北京)科技有限公司

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