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Position: Discussion forums > Bioscience biotechniques discussion forums > Proteomics and protein biochemistry forum
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protein solubility
Posted by: catzzh (IP Hidden, New member, 4)
Date: May 2, 2005 12:36PM
I'm using a protein to treat the K562 cells. The working concentration of the protein is 100uM, but the highest concentration in phosphate buffer+0.1%DMSO is 100uM(initial concentration). If the solution can be used to treat cells, the initial should be 1mM(10 times diluted with medium to treat cells).
Is there any way of improving the protein solubility? Like using some kind of non-toxic solubilizer? Thank you very much!
protein solubility
Posted by: Nilkanth Faldu (IP Hidden, Unregistered user, )
Date: November 13, 2005 05:37AM
Respected sir
Can you Kindly send me the articles on "invivo approach for protein solubilization of overexpressed protein" Regards Nilkanth
Re: protein solubility
Posted by: hazariv (IP Hidden, New member, 2)
Date: December 18, 2005 07:30PM
Try adding a non ionic detergent like tween (0.05 - 0.1 %) should help.
'can also try to keep the pH atleast 1.5 - 2 unots away from the proteins pI.
Re: protein solubility
Posted by: Sciuser (IP Hidden, New member, 2)
Date: December 28, 2005 07:24PM
Hi there. One possible explanation for the poor solubility is that the protein that you are using for your studies is expressed from bacteria (eg. E. coli). The potential limitation of this is that many bacterially expressed proteins are poorly soluble. The reason for this is that bacteria don't add carbohydrate structures. Nature's way of improving solubility of proteins is to glycosylate. Structures such as sialic acids are highly charged and hydrophilic, and completely change the net charge (and aqueous solubility) of the protein. I know apollo cytokine research do contract expression of human cell expressed proteins. Human cell expression yields proteins with the carbohydrate structures and are therefore more soluble in biological fluids. If you want to stick to a detergent type approach, then a very bio-compatible detergent is deoxycholate - this is bile acid and is produced by the human body in large quantities.
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