Starch Assay Kit

Catalog No. ABIN1000285

Product Details

Target: Starch (GLGA)

Application: Biochemical Assay (BCA)

Specificity: 2 μg/mL

Characteristics: Use as little as 10 µL samples. Linear detection range: 2 to 200 µg/mL starch for colorimetric assays and 0.2 to 20 µg/mL for fluorimetric assays.

Components: Assay Buffer: 12 mL. Enzyme A: 120 µL. Enzyme B: 120 µL. Dye Reagent: 120 µL. Standard: 50 µL 50 mg/mL.

Material not included: Pipeting devices, centrifuge tubes, clear flat bottom 96-well plates and plate reader.

Application Details

Comment:

1. This assay is based on a kinetic reaction, the use of a multi- channel pipettor for adding the working reagent is recommended.
2. Interference. Interference. SH-group containing reagents (e.g., DTT, -mercaptoethanol) may interfere with this assay and should be avoided in sample preparation.

Protocol: 1. Equilibrate all components to room temperature. During experiment, keep thawed enzymes in a refrigerator or on ice.
2. Standards and samples: Dilute standard by mixing 5 µL Standard with 1.245 mL dH2O to give 200 µg/mL standard. Dilute standard in dH2O. Transfer 10 µL standard and samples into separate wells of a clear flat-bottom microplate.
3. Working Reagent. For each reaction well, mix 90 µL Assay Buffer, 1 µL Enzyme A, 1 µL Enzyme B and 1 µL Dye Reagent in a clean tube. Transfer 90 µL Working Reagent into each reaction well. Tap plate to mix.
4. Incubate 30 min at room temperature. Read optical density at 570nm (550-585nm). FLUORIMETRIC

Calculation of Results:

Subtract Blank reading (OD570nm or fluorescence intensity) from the standard reading values and plot the OD or F against standard concentrations.

Restrictions: For Research Use only

Handling

Storage: -20 °C

References

Tsuge, Tateno, Sasaki, Hasunuma, Tanaka, Kondo: "Direct production of organic acids from starch by cell surface-engineered Corynebacterium glutamicum in anaerobic conditions." in: AMB Express, Vol. 3, Issue 1, pp. 72, (2014) (PubMed).

Theerawitaya, Boriboonkaset, Cha-Um, Supaibulwatana, Kirdmanee: "Transcriptional regulations of the genes of starch metabolism and physiological changes in response to salt stress rice (Oryza sativa L.) seedlings." in: Physiology and molecular biology of plants : an international journal of functional plant biology, Vol. 18, Issue 3, pp. 197-208, (2013) (PubMed).

Zebeli, Terrill, Mazzolari, Dunn, Yang, Ametaj: "Intraruminal administration of Megasphaera elsdenii modulated rumen fermentation profile in mid-lactation dairy cows." in: The Journal of dairy research, Vol. 79, Issue 1, pp. 16-25, (2012) (PubMed).

Gardner, Lohman, Gerlach, Cooksey, Peyton: "Comparison of CO(2) and bicarbonate as inorganic carbon sources for triacylglycerol and starch accumulation in Chlamydomonas reinhardtii." in: Biotechnology and bioengineering, Vol. 110, Issue 1, pp. 87-96, (2012) (PubMed).

Iqbal, Zebeli, Mazzolari, Bertoni, Dunn, Yang, Ametaj: "Feeding barley grain steeped in lactic acid modulates rumen fermentation patterns and increases milk fat content in dairy cows." in: Journal of dairy science, Vol. 92, Issue 12, pp. 6023-32, (2009) (PubMed).

Target Details

Target: Starch (GLGA)

Alternative Name: Starch (GLGA Products)

Background: Quantitative determination of starch by colorimetric (570nm) or fluorometric (585/530nm) methods.
Procedure: 30 min.

Starch, chemical formula (C6H10O5)n, is a polysaccharide carbohydrate consisting of a large number of glucose units joined together by glycosidic bonds. All plant seeds and tubers contain starch present in the form of amylose and amylopectin. Starch is the most consumed polysaccharide in the human diet. Some starches are digested very quickly, and cause a rapid and large rise in blood sugar. Others are digested more slowly, and some starch, called resistant starch, is not digested in the small intestine at all, and thus causes little or no blood sugar rise. Simple, direct and automation-ready procedures for measuring starch concentrations find wide applications in research and drug discovery. This starch uses a single Working Reagent that combines the enzymatic break down of starch and the detection of glucose in one step. The color intensity of the reaction product at 570nm or fluorescence intensity at gamma em/ex = 585/530nm is directly proportional to the starch concentration in the sample. This simple convenient assay is carried out at room temperature and takes only 30 min.