3-Hydroxybenzoic Acid as an Internal Standard for the High-Pressure Liquid Chromatography Quantitation of Salicylic Acid in Plants

✍ Scribed by Steven M. Eshita

• Publisher: Elsevier Science
• Year: 2001
• Tongue: English
• Weight: 72 KB
• Volume: 289
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.2000.4962

✦ Synopsis

conditions. The membrane was then probed with guinea pig anti-keratin, and the immunoreactive bands were visualized with alkaline phosphatase-conjugated, goat anti-guinea pig antibody using standard protocols. The results, presented in Fig. 1, clearly demonstrate that keratin is present only in the lane in which the Laemmli sample buffer was prepared with unfiltered 2-ME (lane 3). No keratin was found in the lanes containing Laemmli sample buffer prepared with filtered 2-ME (lane 2) or prepared in its absence (lane 1). Hence, the source of keratin is 2-ME and this contaminant can be eliminated by passing the reducing agent through an inert filter with a nominal 3000 molecular weight cutoff.

We hope that this simple and effective means of solving the problem of keratin contamination in gel electrophoresis will be welcomed by both biochemists and manufacturers of reducing agents for the life sciences community.

Acknowledgments.