α amylase assay kit

Manufactured by Solarbio

Sourced in China

The α-amylase Assay Kit is a laboratory tool used to measure the activity of the enzyme α-amylase. α-amylase is an enzyme that catalyzes the hydrolysis of starch into smaller carbohydrate molecules.

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Lab products found in correlation

Starch content assay kit, by Solarbio (2 mentions) Plant soluble sugar content assay kit, by Solarbio (1 mentions) Hydrogen peroxide assay kit, by Solarbio (1 mentions) Total protein assay kit, by Nanjing Jiancheng (1 mentions)

4 protocols using α amylase assay kit

Carbohydrate Dynamics in Crop Development

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The α-amylase Assay Kit (Solarbio, #BC4570, China) was used to test for α-amylase activity, according to the manufacturer's instructions. The starch content was measured with the Starch Content Assay Kit (Solarbio, #BC0700, China). Soluble sugar was separated from starch using 80% ethanol. The acid hydrolysis method was then used to decompose starch into glucose, and the anthrone colorimetric method was used to determine glucose levels, which were then converted to starch content. Quantitative analyses of carbohydrates were performed using the Plant Soluble Sugar Content Assay Kit (Solarbio, #BC0035, China). The reducing ratio (%) of starch content between the heading and the harvest was calculated using the following formula:
The same calculation method was applied to the reducing ratio of soluble sugar concentration from heading to harvest.

Ouyang N., Sun X., Tan Y., Sun Z., Yu D., Liu H., Liu C., Liu L., Jin L., Zhao B., Yuan D, & Duan M. (2021). Senescence-Specific Expression of RAmy1A Accelerates Non-structural Carbohydrate Remobilization and Grain Filling in Rice (Oryza sativa L.). Frontiers in Plant Science, 12, 647574.

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Mutant Strain mut80 Enzyme Assay

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According to the results of the screening, the mutant strain mut80 exhibited higher protein and starch hydrolysis capacity than the other mutants; therefore, it was selected for subsequent research. After the culture was activated, the single colonies of XS-4 and mut80 were transferred to 100 mL NB liquid medium at 37 °C, 200 rpm, and incubated for 3 days. After incubation, the supernatant was collected via centrifugation for enzyme activity testing. The acidic protease and α-amylase activities were measured using a Micro Acidic Proteinase Assay Kit and an α-amylase Assay Kit (Solar Bio, Beijing, China). We followed the kit instructions for operation. Furthermore, in order to determine the genetic stability of mut80, it was continuously subcultured for 20 generations. The activities of acidic protease and α-amylase were measured every 5 generations [23 (link)].

Zhang A., Ma Y., Deng Y., Zhou Z., Cao Y., Yang B., Bai J, & Sun Q. (2023). Enhancing Protease and Amylase Activities in Bacillus licheniformis XS-4 for Traditional Soy Sauce Fermentation Using ARTP Mutagenesis. Foods, 12(12), 2381.

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Biochemical Indicator Quantification Protocols

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Biochemical indicators were measured using corresponding assay kits from Solarbio Life Sciences (Beijing, China) following the manufacturer’s protocols. In detail, the Starch Content Assay Kit (Solarbio, Cat#BC0700), α-Amylase Assay Kit (Solarbio, Cat#BC0615), Hydrogen Peroxide Assay Kit (Solarbio, Cat#BC3595), and Proline Content Assay Kit (Solarbio, Cat#BC0290) were obtained for subsequent spectrophotometric measurements.

Chen M.X., Zhu F.Y., Wang F.Z., Ye N.H., Gao B., Chen X., Zhao S.S., Fan T., Cao Y.Y., Liu T.Y., Su Z.Z., Xie L.J., Hu Q.J., Wu H.J., Xiao S., Zhang J, & Liu Y.G. (2018). Alternative splicing and translation play important roles in hypoxic germination in rice. Journal of Experimental Botany, 70(3), 817-833.

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Recombinant Amylase and Cellulase Production

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The amylase genes amyE1 (lacking the N-terminal sequence encoding the signal peptide, LNSP) and celE1 (LNSP) were cloned into the pET28a vector and then were transferred into E. coli DE3 cells. The transformants of DE3 were incubated in the liquid LB medium (100 μg/L of ampicillin) at 37°C to an OD₆₀₀ value of about 0.6–0.8 and were transferred into an incubator supplemented with IPTG (0.01%, w/v) at 16°C overnight to induce gene expression. The harvested cells were decomposed and homogenized by sonication (Scientz-II D, Ningbo). The recombinant proteins were purified by using Ni-NTA columns according to the previous reported protocol (Dai et al., 2019 (link)). The activities of amylase and cellulase were assayed by using Cellulase (CL) Assay Kit and α-amylase Assay Kit (Beijing Solarbio Science & Technology Co., Ltd). One unit (U) of enzyme activity was quantified as the amount of enzyme that synthesized against 1 μmol of glucose per minute under the assay conditions. The enzyme concentrations were measured by using a total protein assay kit (Jiancheng Biotech, Nanjing, China). Primers used in this study are listed in Supplementary Table S1.

Dai J., Dong A., Xiong G., Liu Y., Hossain M.S., Liu S., Gao N., Li S., Wang J, & Qiu D. (2020). Production of Highly Active Extracellular Amylase and Cellulase From Bacillus subtilis ZIM3 and a Recombinant Strain With a Potential Application in Tobacco Fermentation. Frontiers in Microbiology, 11, 1539.

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