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This protocol quantifies cysteine residues from small amounts of pollen (10 mg) after phenylisothiocyanate (PITC) derivatization (adapted from Manneberg et al 1995). Quantification of cysteine by other methods is difficult because of its instability during amino acid analysis. Samples of anther pollen, corbicular pollen, or stored pollen from bee colonies are completely digested in strong acid. Cysteine residues are completely oxidized to cysteic acid then derivatized with PITC before HPLC UV VIS analysis. Manneberg, M., Lahm, H., & Fountoulakis, M. (1995). Quantification of cysteine residues following oxidation to cysteic acid in the presence of sodium azide. Analytical Biochemistry, 231, 349-353. https://doi.org/10.1006/abio.1995.9988 National Program Number: NP 305 Project Number: 2022-21000-022-000D, 2022-21000-022-053-I Carl Hayden Bee Research Center, USDA-ARS PWA, Tucson, Arizona, USA Grand Challenge - Assessing the Nutrient Contents of Pollen for Bees |
