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A method for the determination of conifer foliage protein extracted using sodium dodecyl sulphate and mercaptoethanol
Author(s) -
Ekramoddoullah K. M.,
Davidson Joanne J.
Publication year - 1995
Publication title -
phytochemical analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.574
H-Index - 72
eISSN - 1099-1565
pISSN - 0958-0344
DOI - 10.1002/pca.2800060103
Subject(s) - chemistry , chromatography , sodium dodecyl sulfate , bradford protein assay , extraction (chemistry) , protein purification , sodium , coomassie brilliant blue , membrane , staining , biochemistry , organic chemistry , medicine , pathology
Sodium dodecyl sulphate and mercaptoethanol are commonly used for extraction and solubilization of proteins, but these substances interfere with previously reported methods for the quantitation of small amounts of protein. In order to overcome this difficulty, a method has been developed for assaying small quantities of protein by (i) spotting the protein solution on a polyvinylidene difluoride membrane, (ii) staining the membrane bound protein with Coomassie Blue, (iii) destaining, and (iv) determining the optical density of the stained membrane by laser‐based scanning. The determination of proteins in conifer foliage by this method is described.