Glucose Assay Kit
中文名称 : 葡萄糖检测分析试剂盒
葡萄糖是自然界分布最广且最为重要的一种单糖,它是一种多羟基醛。葡萄糖水溶液旋光向右,故亦称“右旋糖”。葡萄糖在生物学领域具有重要地位,是活细胞的 能量来源和新陈代谢中间产物。植物可通过光合作用产生葡萄糖。在糖果制造业和医药领域有着广泛应用。葡萄糖是细胞中重要的能量来源,能产生普遍的能量分子 ATP。血糖水平是诊断多种代谢紊乱的关键参数之一。
葡萄糖检测试剂盒能够快捷的检测各种生物样品(比如血清、血浆、其它体液、食品、生长培养基等等)中葡萄糖的含量。检测实验中,葡萄糖被特异性氧化,产物 与一种染料反应显色(λ = 570 nm)和荧光(Ex/Em = 535/587 nm)。其吸光度及荧光强度与葡萄糖含量成正比例关系。本检测技术快速简单、敏感性高,适应于高通量分析。该葡萄糖检测试剂盒也适用于监测发酵过程中或葡 萄糖依赖型蛋白质表达过程中的葡萄糖含量。检测量程为1-10000M。
1、步骤简单,只需30分钟;2、操作快速并且方便
Glucose Assay Kit
(Catalog #K606-100; 100 assays; Store at -20 °C.)
I. Introduction:
Glucose (C6H12O6; FW: 180.16) is a very important fuel source to generate the universal
energy molecule ATP. Glucose level is a key diagnostic parameter for many metabolic
disorders. Measurement of glucose can be very important in both research and drug
discovery processes. The BioVision Glucose Assay Kit provides direct measurement of
glucose in various biological samples (e.g., serum, plasma, other body fluids, food, growth
media, etc.). Glucose Enzyme Mix specifically oxidizes glucose to generate a product
which reacts with a dye to generate color (λ = 570 nm) and fluorescence (Ex/Em =
535/587 nm). The color and fluorescence generated is proportional to the amount of
glucose present. The method is rapid, simple, sensitive, and suitable for high throughput.
The assay is also suitable for monitoring glucose level during fermentation and glucose
feeding in protein expression processes. The kit detects 1-10000 μM glucose samples.
II. Kit Contents
III. Storage and Handling:
Store kit at –20°C, protect from light. Warm the Glucose Assay Buffer to room temperature
and briefly centrifuge vials before opening. Read the entire protocol before performing the
assay.
IV. Reagent Preparation:
Glucose Probe: Ready to use as supplied. Warm to > 18 °C (~room temp) prior to
use to melt frozen DMSO. Store at –20 °C, protect from light and
moisture. Use within two months.
Glucose Enzyme Mix: Dissolve in 220 μl Glucose Assay Buffer, then aliquot & store at
–20 °C. Keep on ice while in use. Use within two months.
V. Glucose Assay Protocol:
1. Standard Curve Preparations:
For colorimetric assay,dilute the Glucose Standard to 1 nmol/μl by adding 10 μl of the
Glucose Standard to 990 μl of Glucose Assay Buffer, mix well. Add 0, 2, 4, 6, 8, 10 μl into
a series of well on a 96 well plate. Adjust volume to 50 μl/well with Glucose Assay Buffer
to generate 0, 2, 4, 6, 8, 10 nmol/well of Glucose Standard.
For the fluorometric assay, dilute the Glucose Standard solution to 0.1 nmol/μl by adding
10 μl of the Glucose Standard to 990 μl of Glucose Assay Buffer, mix well. Then take 20 μl
into180 μl of Glucose Assay Buffer. Mix well. Add 0, 2, 4, 6, 8, 10 μl into a series of wells
as in the colorimetric assay. Adjust volume to 50 μl/well with Glucose Assay Buffer to
generate 0, 0.2, 0.4, 0.6, 0.8, 1.0 nmol/well of the Glucose Standard.
2. Sample Preparation: Prepare test samples at a total volume of 50 μl/well with Glucose
Assay Buffer in a 96-well plate. If using serum, limit sample volume to 0.5-2 μl/assay. Adjust
the final volume to 50 μl with Assay buffer. Normal serum contains ~5 nmol/μl glucose can
be directly diluted in the Glucose Assay Buffer. We suggest testing several doses of your
sample to make sure the readings are within the standard curve range.
3. Glucose Reaction Mix: Mix enough reagent for the number of assays to be performed: For
each well, prepare a total 50 μl Reaction Mix containing:
46 μl Glucose Assay Buffer
2 μl Glucose Probe*
2 μl Glucose Enzyme Mix
*Note: The fluorometric assay is ~10 times more sensitive than the colorimetric assay. Use
0.4 μl of the probe per reaction to decrease the background reading / increase detection
sensitivity significantly.
4. Mix well. Add 50 μl of the Reaction Mix to each well containing the Glucose Standard and
test samples. Mix well.
5. Incubate the reaction for 30 minutes at 37 °C, protect from light. Measure absorbance at 570
nm for colorimetric assay or Ex/Em = 535/590 nm for fluorometric assay in a microplate
reader.
6. Calculations: Correct background by subtracting the value derived from the 0 glucose
control from all readings (Note: The background reading can be significant and must be
subtracted from sample readings). Glucose concentrations of the test samples can then be
calculated.
C = Sa/Sv (nmol/μl or μmol/ml, or mM)
Where Sa is sample amount (in nmol) from standard curve.
Sv is sample volume (in μl) added into the sample wells.
Glucose Molecular Weight 180.16.
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Component K606-100 Cap Code Part Number
Glucose Assay Buffer 25 ml WM K606-100-1
Glucose Probe (in DMSO) 0.2 ml Red K606-100-2A
Glucose Enzyme Mix (lyophilized) 1 vial Green K606-100-4
Glucose Standard (100 nmol/l) 100 μl Yellow K606-100-5
y = 0.128x + 0.010
0
0.5
1
1.5
0 2 4 6