Greetings all,
I am purifying protein from the squid ovary using ammonium sulfate precipitation (ASP). After centrifugation, I am getting a gel like pellet (not exactly pellet) which is easy to soluble in the extraction buffer (Phosphate buffer 10mM, pH 7, containing NaCl; 0.45M). I have increased centrifugation force and time but still results are same.
Squid ovary contained around 27% (dry base) of carbohydrates. Is it possible that carbohydrates are hindering in this purification step?
Before ASP step I did extraction of desired protein using extraction buffer (mentioned above) and then centrifuge at 10,000 xg and got solid pellet and then supernatant was subjected to ASP.
Thanks for any help on this,
Regards
Avtar
Thank you for your answer Dear Schaefer.
Chemical composition is affecting the exaction procedure. Therefore, I am thinking to remove carbohydrates from squid ovary before purification.
For Protein content it is (67%) the first major component of squid ovary so I think, protein content is not a problem.
Dear Schaefer,
Thank you so much for your valuable suggestions. I will follow what you have suggested.
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