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This protocol quantifies total protein contents of small amounts of pollen (10 mg) using the bicinchoninic acid (BCA) assay of the Pierce BCA protein assay kit (Thermo Scientific product number 23225). This protocol adapts the kit protocol for analysis of anther pollen, corbicular pollen, and stored pollen from bee colonies. Pollen samples are subjected to acid hydrolysis to digest refractory proteins and to degrade sugars and other pollen substances that interfere with total protein assays. The acid hydrolysis releases substantially more protein from pollens than occurs with soluble proteins but may result in some protein losses due to the harsh acidic conditions. Similar BCA analysis methods without hydrolysis may be used to quantify soluble proteins (proteins obtained without acid hydrolysis or other digest methods) which may exclude insoluble pollen proteins from full quantification. The total protein contents obtained using BCA analysis of acid hydrosylates represents the maximum protein available to consumers and likely includes some refractory proteins that may not be digested by the consumer. The assay is modified from the original standard protocol to work for pollen analysis: Document Connect (thermofisher.com). Please note that our steps differ from the commercial source protocol. 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 |
