• »
• Protein extraction from marine phytoplankton

Protein extraction from marine phytoplankton

Instrument: Precellys® 24 Dual

Sample: Culture pellets from Ostreococcus tauri, Thalassiosira pseudonana, Synechococcus sp. WH7803, Prochlorococcus marinus PCC 9511 and Synechococcus sp. WH8101 re-suspended in buffer.

Protocol:

• Sample suspension consists of culture pellets from Ostreococcus tauri, Thalassiosira pseudonana, Synechococcus sp. WH7803, Prochlorococcus marinus PCC 9511 and Synechococcus sp. WH8101 re-suspended in Protein extraction buffer (Agrisera, Cat. AS08 300) supplemented with the protease inhibitor 4-(2-aminoethyl)-benzene-sulfonyl fluoride HCl (417μM).
• Add 400μl of suspension to 2mL Precellys tube
• Run the Precellys at 6500 rpm, 3x20 sec (5 sec break) with Cryolys.

Results: Total protein quantification, Immunoblotting with relevant antibodies. Sample throughput and reproducibility are higher than with sonication/flash freeze protocol

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 and a horseradish peroxidase-conjugated goat anti-chicken secondary antibody.