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  • Evaluation of extraction solutions for biochemical analyses of the proteins in rice grains.

Evaluation of extraction solutions for biochemical analyses of the proteins in rice grains.

Bioscience, biotechnology, and biochemistry (2013-01-08)
Gang-hua Lang, Yukari Kagiya, Mayumi Ohnishi-Kameyama, Kazumi Kitta
ABSTRACT

The influence of different extraction solutions on the proteins extracted from rice grains was investigated. The largest amounts of salt-soluble proteins were extracted with solutions supplemented with Tris-HCl at pH 8.0. Rice allergens were analyzed by multiplex immunodetection. Except for α-globulin extracted with the solutions at pH 8.0, which showed a low-molecular-weight band besides the main band, no significant solution-dependent difference among the allergens was found. Total proteins were extracted with four kinds of solution. The extraction of the basic subunit of glutelin was found to be SDS-dependent, and more protein was obtained with extraction solutions supplemented with SDS. The contents of α-globulin and α-amylase/trypsin inhibitors were higher in the extracts without SDS than with SDS. We conclude from the present data that, in order to obtain comparable data from rice grain salt-soluble and total protein analyses, differences in the protein extraction efficiency of solutions used should be taken into consideration.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
α-Amylase from human saliva, Type XIII-A, lyophilized powder, 300-1,500 units/mg protein
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α-Amylase from Aspergillus oryzae, aqueous solution, ≥800 FAU/g
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α-Amylase from Bacillus amyloliquefaciens, liquid, ≥250 units/g
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α-Amylase from Bacillus licheniformis, Type XII-A, saline solution, ≥500 units/mg protein (biuret)
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α-Amylase from Bacillus licheniformis, suitable for determination of starch (Kit STA-20)
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α-Amylase from human saliva, Type IX-A, lyophilized powder, 1,000-3,000 units/mg protein
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α-Amylase from Bacillus licheniformis, lyophilized powder, 500-1,500 units/mg protein, 93-100% (SDS-PAGE)
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α-Amylase from Aspergillus oryzae, powder, ~30 U/mg
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α-Amylase from Aspergillus oryzae, ≥150 units/mg protein (biuret)
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α-Amylase from Aspergillus oryzae, powder, ~1.5 U/mg (~0.2 U acc. to Willstätter)
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α-Amylase from Bacillus sp., liquid
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α-Amylase from Bacillus sp., powder, ≥400 units/mg protein (Lowry)
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α-Amylase from porcine pancreas, PMSF Treated, Type I-A, saline suspension, ≥1000 units/mg protein (E1%/280)
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α-Amylase, heat-stable, solution, for use in Total Dietary Fiber Assay, TDF-100A
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α-Amylase from Bacillus sp., Type II-A, lyophilized powder, ≥1,500 units/mg protein (biuret)
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α-Amylase from porcine pancreas, Type VI-B, ≥5 units/mg solid
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α-Amylase from Bacillus sp., powder, yellow-brown, ~380 U/mg
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α-Amylase from Bacillus sp., powder, yellow-brown, ~50 U/mg
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α-Amylase from porcine pancreas, Type I-A, PMSF treated, saline suspension, 700-1400 units/mg protein (E1%/280)
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Taka-Diastase from Aspergillus oryzae, powder, slightly beige, ~100 U/mg