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Steven Burgess 2020. Leaf Protein Extraction for Immunoblot (Soybean, Cowpea, Tobacco). protocols.io dx.doi.org/10.17504/protocols.io.bqnmmvc6Copy Citation Copied
URL: https://dx.doi.org/10.17504/protocols.io.bqnmmvc6
Authors: Steven Burgess
Group: Ag SynBio Lab UIUC
Summary: This protein extraction protocol was developed for analysis of protein abundance in leaf tissue by immunoblot. It was optimized for dicot species including Glycine max.Quantities - 450 µL protein extraction buffer (PEB) per three 13.4 mm diameter leaf disks (size #7 Humboldt Cork Borer)Note from Agrisera: Keeping sample volumes in a range of 0.2-0.5 mL has been found to contribute to better extraction results, an upscale in volume is not recommended, if no cork borer available the suggestion is 500 µL PEB per 100 mg of plant tissue. - Final volume ~400 µL - Expected yield: 1.5 - 6 µg / µL - Total yield: 6 - 24 mg protein - Amount of total soluble protein to load per lane 30 µg Note: when using the TissueLyser II it is recommended to use 2 mL centrifuge tubes in conjunction with 4mm beads. 1.5mL tubes are narrow at the end and samples will not grind properly if you use the 4mm beads.
Affiliations: University of Illinois at Urbana-Champaign
Version: 2
Publication Date: 2020
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Source: Protocols.io
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