The L-Rhamnose Assay Kit for the measurement of L-rhamnose in plant extracts, culture media/supernatants and other materials is a simple, rapid method.
L-Rhamnose occurs naturally in the L-form and is commonly present as a component of the carbohydrate moiety of eukaryotic glycoproteins and in plant cell wall polysaccharides. The most abundant occurrence of L-rhamnose is within the pectic fraction of plant cell wall polysaccharides. L-Rhamnose is commonly used as a non-metabolisable marker along with lactulose for dual-permeability testing in the diagnosis of intestinal diseases such as Crohn’s disease or coeliac disease.
Note for Content: The number of manual tests per kit can be doubled if all volumes are halved. This can be readily accommodated using the MegaQuantTM Wave Spectrophotometer (D-MQWAVE).
View more of our monosaccharide and disaccharide assay kits.
Content: 50 / 100 assays (manual) / 550 assays (microplate) / 550 assays (auto-analyser)
Shipping Temperature: Ambient
Storage Temperature: Short term stability: 2-8oC,
Long term stability: See individual component labels
Stability: > 1 year under recommended storage conditions
Analyte: L-Rhamnose
Assay Format: Spectrophotometer, Microplate, Auto-analyser
Detection Method: Absorbance
Wavelength (nm): 340
Signal Response: Increase
Linear Range: 5 to 100 µg of L-rhamnose per assay
Limit of Detection: ~ 1.2 mg/L
Reaction Time (min): ~ 5 min at 25oC or ~ 4 min at 37oC
Application examples: Hydrolysates of plant material and polysaccharides, culture media / supernatants and other materials.
Method recognition: Novel method
Advantages
Very cost effective
All reagents stable for > 2 years after preparation
Only test kit available
Simple format
Rapid reaction (~ 5 min at 25oC or ~ 4 min at 37oC)
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
用于测量植物提取物、培养基/上清液和其他材料中L-鼠李糖的L-鼠李糖含量测定试剂盒是一种简单、快速的方法。
L-鼠李糖以L-形式自然存在,通常作为真核糖蛋白的碳水化合物部分和植物细胞壁多糖的成分存在。L-鼠李糖最丰富的存在是在植物细胞壁多糖的果胶部分中。L-鼠李糖通常与乳果糖一起用作非代谢标记物,用于诊断克罗恩病或腹腔疾病等肠道疾病的双重渗透性测试。
内容说明:如果所有试剂盒的体积减半,每个试剂盒的手动测试数量可以翻倍。这可以使用MegaQuantTM波分光光度计(D-MQWAVE)轻松调节。
查看更多我们的单糖和双糖检测试剂盒。
含量:50/100测定(手动)/550测定(微孔板)/550检测(自动分析仪)
运输温度:环境温度
储存温度:短期稳定性:2-8oC,
长期稳定性:参见单个组件标签
稳定性:在推荐的储存条件下>1年
分析物:L-鼠李糖
测定形式:分光光度计、微孔板、自动分析仪
检测方法:吸光度
波长(nm):340
信号响应:增加
线性范围:每次测定5至100µg L-鼠李糖
检测限:~1.2 mg/L
反应时间(分钟):在25℃时约5分钟或在37℃时约4分钟
应用实例:植物材料和多糖的水解物、培养基/上清液和其他材料。
方法识别:一种新颖的方法
优点
非常经济高效
所有试剂在制备后稳定超过2年
仅提供检测试剂盒
简单格式
快速反应(在25℃下约5分钟或在37℃下约4分钟)
Mega Calc™我们的网站上提供了软件工具,可以轻松处理原始数据
包括标准
适用于手动、微板和自动分析仪格式
Q1. 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.
Sometimes the addition of the last assay component can cause a small negative absorbance change in the blank samples due to a dilution effect and in such cases it is recommended that the real absorbance values be used in the calculation of results.
If you suspect that the Megazyme test kit is not performing as expected such that expected results are not obtained please do the following:
For samples with low concentrations of analyte the sample volume used in the kit assay can be increased to increase sensitivity. When doing this the water volume is adjusted to retain the same final assay volume. This is critical for the manual assay format because the assay volume and sample volume are used in the calculation of results.
The volume/weight of sample and total volume of the extract can be modified to suit the sample. This will ultimately be dictated by the amount of analyte of interest in the sample and may require empirical determination. For low levels of analyte the sample:extract volume ratio can be increased (i.e. increase the sample and/or decrease the total extraction volume).
Alternatively, for samples with low concentrations of analyte, a larger sample volume can be added to the kit assay. When altering the sample volume adjust the distilled water volume added to the assay accordingly so that the total assay volume is not altered.
The kit assay may work for biological fluids assuming that inositol is present above the limit of detection for the kit after any sample preparation (if required). Centrifugation of the samples and use of the supernatant directly in the kit assay (with appropriate dilution in distilled water) may be sufficient. However, if required a more stringent sample preparation method may be required and examples are provided at the following link:http://www.megazyme.com/docs/analytical-applications-downloads/biological_samples_111109.pdf?sfvrsn=2
The test kit has not been tested using biological fluids as samples because it is not marketed or registered as a medical device. This will therefore require your own validation.
The majority of the Megazyme test kits are developed to work in cuvettes using the manual assay format, however the assay can be converted for use in a 96-well microplate format. To do this the assay volumes for the manual cuvette format are reduced by 10-fold. The calculation of results for the manual assay format uses a 1 cm path-length, however the path-length in the microplate is not 1 cm and therefore the MegaCalc spreadsheet or the calculation provided in the kit booklet for the manual format cannot be used for the micropalate format unless the microplate reader being used can.
There a 3 main methods for calculation of results using the microplate format:
Where the amount of analyte in a liquid sample is unknown, it is recommended that a range of sample dilutions are prepared with the aim of obtaining an absorbance change in the assay that is within the linear range.
Where solid samples are analysed, the weight of sample per volume of water used for sample extraction/preparation can be altered to suit, as can the dilution of the extracted sample prior to the addition of the assay, as per liquid samples.
If there are any concerns with any kit components, the first thing to do is to test the standard sample (control sample) that is supplied with the kit and ensure that the expected value (within the accepted variation) is obtained before testing any precious samples. This must be done using the procedure provided in the kit booklet without any modifications to the procedure. If there are still doubts about the results using the standard sample in the kit then send example results in the MegaCalc spread sheet to your product supplier (Megazyme or your local Megazyme distributor).
The kit assay will only measure the non-covalently linked monosaccharide.
Oligosaccharides or polysaccharides can be measured after hydrolysis to the monosaccharide. Generally acid hydrolysis can be achieved by boiling the oligo/polysaccharide in 1.3 M HCl for 1 h. It is recommended that scientific literature is consulted for information on hydrolysis conditions for the particular oligo/polysaccharide that is being measured.
No. The 0.1 change of absorbance is only a recommendation. The lowest acceptable change in absorbance can is dictated by the analyst and equipment (i.e. pipettes and spectrophotometer) and therefore can be can be determined by the user. With accurate pipetting, absorbance changes as low as 0.02 can be used accurately.
If a change in absorbance above 0.1 is required but cannot be achieved due to low concentrations of analyte in a sample, this can be overcome by using a larger sample volume in the assay to increase the absorbance change and thereby increase sensitivity of the assay. When doing this the increased volume of the sample should be subtracted from the distilled water volume that is added to the assay so that the total assay volume is unaltered. The increase sample volume should also be accounted for when calculating final results.
Yes. Samples with the lower concentrations of analyte will generate a lower absorbance change. For samples with low concentrations of analyte, a larger sample volume can be used in the assay to increase the absorbance change and thereby increase sensitivity of the assay. When doing this the increased volume of the sample should be subtracted from the distilled water volume that is added to the assay so that the total assay volume is unaltered. The increase sample volume should also be accounted for when calculating final results.
The test kit is extremely accurate – at Megazyme the quality control criteria for accuracy and repeatability is to be within 2% of the expected value using pure analytes.
However, the level of accuracy is obviously analyst and sample dependent.
Yes, instead of adding 2 μL of enzyme suspension an alternative is to dilute the enzyme and add a larger volume to the microplate assay.
Dilute the assay buffer 10-fold with distilled water and use this as the diluent to dilute an aliquot of the enzyme suspension also by 10-fold. Instead of 2 μL, use 20 μL of the diluted enzyme in the microplate assay.
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