Hi there,

Protein aggregates are usually not soluble which means they are easily pelleted with cell debris by centrifugation.

Unless you want the protein to be in the form of aggregates, it would be worthwhile to work on finding expression conditions that result in unaggregated protein.

Hi,

Protein aggregation could be a result any of the following factors, to start most importantly mindful of the "amino acid sequence" if the aa's have too many hydrophobic regions this could lead to protein aggregation. Post translational modifications can do that too, and depends if you are expressing a prokaryote/eurkaryote gene and the type of host expression system.

Also, try to check your expression conditions, pH, temperture (low temp is best: highly recomended ), Concentration of the protein itself can lead to aggregation.

Presence of denaturing agents like DDT Urea can also lead to this.

For Lysis buffer try the combination of Bugbuster + Benzonase + rLysozyme (depends on your expression vector) + EDTA free protease inhibitor cocktails.

Good luck

Hi Arvind ,

thanks for the answer but the goal is to keep my aggregates and only separate them , i need them separated from the rest of the lysate .