- Azo-果聚糖
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英文名:Azo-Fructan – 5gr
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货号:S-AZFR5
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规格:5 grams
市场价: 3300元
Substrate for the measurement of endo-inulinase.
内切型菊粉酶检测底物
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英文名:Azo-Fructan – 5gr
货号:S-AZFR5
规格:5 grams
市场价: 3300元
Substrate for the measurement of endo-inulinase.
内切型菊粉酶检测底物
PDF Download
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英文名:Fructan HK Assay Kit
货号:K-FRUCHK
规格:50 assays per kit
市场价: 5194元
分析物意义:许多食品如洋葱和种子中的常见组分
Megazyme检测试剂盒优点:方法新颖、反应快、试剂稳定
The Fructan HK test kit is suitable for the specific measurement and analysis of all fructo-oligosaccharides (reducing and non-reducing) and of fructan polysaccharide.
UV-method for the determination of Fructan in foodstuffs,
beverages and other materials
Principle:
(sucrase + maltase)
(1) Sucrose + maltosaccharides + H2O → D-glucose + D-fructose
(exo-inulinase + endo-inulinase)
(2) Fructan + H2O → D-glucose + D-fructose
(hexokinase)
(3) D-Glucose + D-fructose + ATP → G-6-P + F-6-P + ADP
(glucose-6-phosphate dehydrogenase)
(4) G-6-P + NADP+ → gluconate-6-phosphate + NADPH + H+
(phosphoglucose isomerase)
(5) F-6-P ↔ G-6-P
Kit size: 50 assays
Method: Spectrophotometric at 340 nm
Total assay time: ~ 90 min
Detection limit: 1-100% of sample weight
Application examples:
Flours, plant materials (e.g. onion), food products and other materials
Method recognition:
This method is a modification of AOAC Method 999.03 and AACC
Method 32-32.01
Advantages
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:
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 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:
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 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.
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.
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