I found only papers involving density gradients. This is too difficult for us, because we have no ultracentrifuge
Raghad Mouhamad
Cell Lysis: Harvest cyanobacteria cells, resuspend in a buffer (e.g., Tris-HCl, NaCl), and lyse using sonication, French press, or detergents like DDM or Triton X-100. Clarification: Centrifuge the lysate to remove debris, and collect the supernatant containing the soluble PSI. Affinity Chromatography: Use Ni-NTA affinity chromatography if PSI is His-tagged or blue Sepharose chromatography for untagged PSI. Ion Exchange Chromatography: Apply anion-exchange chromatography (e.g., DEAE-Sepharose or Q-Sepharose) to further purify PSI based on charge. Size Exclusion Chromatography: Use gel filtration (e.g., Sephacryl S-300 or Superdex 200) to separate PSI by size. Detergent Optimization: Use mild detergents like DDM or LMNG to maintain PSI solubility and stability.
To purify Photosystem I (PSI) from cyanobacteria using chromatography instead of sucrose density gradients, follow these steps:
This method allows PSI purification without sucrose density gradients and can be adapted for different purity levels.
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