During the experiment and literature I found that many peoples had used tris Hcl for protein extraion in the range of 20 to 100 mM concentration. why peoples are not extracting proteins at above this concentration ??? I study the various concentration of Tris- HCL for protein extraction from 0.2 M to 2.0 M and I found the highest soluble protein at 1.6 M concentration of Tris HCL. so I want to know that is am going in right way???
No high concentration of tris HCl buffer affect 3 d structure of proteins mainly dislodge its intrachain interactions, though protein do not denature but form abrupt changes in structure. Contrary to this if high concentration of any alkali is used, protein get denature. For maintaining structural integrity of proteins they should need only a pH range between 5.2 to 6.9.
Hi Sachin, every protein extraction procedure will be different depending on what you are trying to extract.
If 1.6 M tris-HCl works for you, then go ahead and use it.
You don't have refer to previous publications about protein extraction using tris-HCl. Just show that the 1.6M tris-HCl works best for you and leave it at that.
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