Agrisera Protein Extraction Buffer PEB and global antibodies (anti-PsbA and anti-RbcL) were used to validate high throughput protein extraction from tough, recalcitrant marine phytoplankton. A standard microprobe sonication/flash freeze protocol was compared with Precellys® (Bertin Technologies) bead-based system to extrat total proteins from multiple samples in parallel. In image: comparison of ribulose-1,5-bisphosphate carboxylase oxygenase (RbcL) immunodetection from phytoplankton, after protein extraction through sonication or Precellys bead beating. RbcL was detected by immunoblotting using a chicken anti-RbcL primary antibody (AS01 017) and a horseradish peroxidase-conjugated goat anti-chicken secondary antibody (AS09 603). MW, molecular weight protein ladder (in kDa). The total yield of proteins extracted from each species was similar using the sonication/flash freeze protocol or Precellys® bead beating system. Using the Precellys® system, sample throughput and reproducibility are higher than with the sonication/flash freeze protocol. |
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