One‐step purification of the β‐glucan elicitor‐binding protein from soybean (Glycine max L.) roots and characterization of an anti‐peptide antiserum

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Abstract
A low abundance β‐glucan elicitor‐binding protein from soybean was isolated by a rapid, simple and one‐step purification method yielding about 9000‐fold enrichment. The affinity‐based purification technique was more efficient than a procedure that uses conventional methods and preserved the binding activity to a much larger extent. The final preparation consisted of one major protein with an apparent molecular mass of about 75 kDa. Electrophoretic analyses of the purified and photoaffinity‐labeled binding protein showed that the native protein was an oligomer with apparent molecular mass of about 240 kDa. A polyclonal anti‐peptide antiserum was raised against a synthetic 15‐mer internal oligopeptide sequence derived from the 75‐kDa protein. The antiserum recognized the purified binding protein in immunoblotting experiments and precipitated the affinity‐labeled protein from a crude extract of the membrane fraction.