Hello all I am working with a small protein-protein complex. This complex express quite well . I purify in a buffer of pH=9.0 with 150mM NaCl and 1% of glycerol and able to concentrate upto 20 mg per ml. I have a two clones of this protein complex. One is N-terminal His tagged and another C-terminal His-tagged. While concentration of the N-terminal His tagged protein in cnetricon it forms yellow color slimy deposition on the surface of membrane. while C-terminal His tagged protein does form very highly viscous layere at the surface of membrane but it is completly colourless.I aliquate the concentrated protein by pippetting into different aliquate rather than collection of whole protein by centrifugation. if i check the concentration of the last aliqoute(which isbottommost viscous protein part) both prtoein complex, it shows very high concentration compared to the first fraction. I have not done DLS. Is my both C-terminal His tagged tagged as well as N-terminal His tagged protein are forming soluble aggregates.
I would appreciate the help. Thanks in advance Peter
