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Starch Colorimetric/Fluorometric Assay Kit

Catalog #: K647
$405.00

Product Details

Cat # +Size K647-100
Size 100 assays
Detection Method Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm)
Species Reactivity Mammalian
Applications The assay can detect starch at 0.0004 to 2 mg/ml.
Features & Benefits • Simple procedure; takes ~ 40 minutes
• Fast and convenient
• Kit contains all necessary reagents for accurate measurement of Starch levels
Kit Components • Hydrolysis Buffer
• Development Buffer
• OxiRed Probe
• DMSO (anhydrous)
• Hydrolysis Enzyme Mix
• Development Enzyme Mix
• Starch Standard (2.0 mg/ml)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Starch is a complex carbohydrate consisting of a large number of glucose units. All plants contain starch, present as amylose, (linear α-1,4 linked polymer) and amylopectin, (highly α-1,6 branched α-1,4 polymer). Starch generally contains 0-25% amylose and 75–100% amylopectin. The Starch Assay Kit provides an easy, accurate assay to measure starch levels in a variety of samples. In the assay, starch is hydrolyzed to glucose which is oxidized to generate color (570 nm) and fluorescence (Ex/Em = 535/587 nm). The assay can detect starch at 0.0004 to 2 mg/ml.


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What is the method to isolate total starch from samples?
For total starch, both the soluble and resistant starch methods (sections V2A and V2B on the datasheet) are used.
What is the expected starch percentage in potato or root vegetable samples?
The percentage varies from one to another root vegetable and even between the types of potato.
What are some tips in case the readings are very low?
1. The buffers should be warmed up to room temperature. Otherwise the reaction will proceed very slowly and can result in such low readings. There is no need to heat samples to 37C.
2. The probe must be at RT and the DMSO must be completely thawed.
3. Ensure that the standard solutions are prepared without pipetting errors.
4. Incubate at RT for 30mins after adding the hydrolyzing enzyme to the standards. Without complete hydrolysis, measurements can be every low.
5. The readings must be taken at 570nm.
For cuvette-based measurements, is it OK to set up the reaction as for the 96-well format and then dilute with just distilled water to make up 500-600 ul and measure?
It is not recommended at all to dilute samples with the reaction mix with water for making up more than 2 times the volume. This will compromise the efficiency of the assay and the measurements made. More importantly for different dilutions of the standard and multiple samples to be compared, the time of incubation should be the same for all. This is not possible to be achieved if you will be using cuvettes. We recommend scaling up all volumes proportionally to do the assay in the volume required for cuvette measurements.
Can individual components of this kit be purchased separately?
Yes, any of the kit's components can be purchased separately without having to buy the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can frozen samples be used with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
Is it essential to make a standard curve for every expt, or is one curve/kit enough?
Yes, it is strongly recommended to do the standards every time you do the expt. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.
I like to use this kit to quantify starch in pet food and lyophilized fecal samples. Is it possible to use this kit for these sample types? Which types of samples are compatible with the assay?
We have not tested pet food or fecal samples. That being said, theoretically you should be able to use it with this kit. A variety of samples should be compatible with the assay. We have tested corn starch, rice starch and wheat starch with this kit.