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.
Standard microprobe sonication/flash freeze protocol has been compared with Precellys® (Bertin Technologies) bead-based system to extract total proteins from multiple samples in parallel.
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 Precellys® system, sample throughput and reproducibility are higher than with the sonication/flash freeze protocol.