The problem is that DMSO has a cut-off absorption below 270nm and the protein shows CD signal in the far-UV region of about 208nm and hence I cannot measure the signal properly because of the high absorption of dmso in that region.
Please suggest me with a solution as to how do i overcome this problem?
You could try to dissolve your compound in the same buffer as the protein, or dissolve it in a different water-miscible solvent that doesn't absorb so much in the far-UV. If that is not possible, the best you could do is to maximize the protein-to-DMSO ratio and minimize the path-length of the cuvette.
Make the stock solution of the compound in DMSO as concentrated as possible. If you already have dissolved all of the solid compound, you can evaporate the DMSO using a lyophilizer (it's slow but it can be done).
By using a high protein concentration, you can decrease the path-length of the cuvette, using the type that consists of two separate pieces with a thin gap between them. By decreasing the path-length, the absorbance due to the solvent will be reduced proportionately.
You could try to dissolve your compound in the same buffer as the protein, or dissolve it in a different water-miscible solvent that doesn't absorb so much in the far-UV. If that is not possible, the best you could do is to maximize the protein-to-DMSO ratio and minimize the path-length of the cuvette.
Make the stock solution of the compound in DMSO as concentrated as possible. If you already have dissolved all of the solid compound, you can evaporate the DMSO using a lyophilizer (it's slow but it can be done).
By using a high protein concentration, you can decrease the path-length of the cuvette, using the type that consists of two separate pieces with a thin gap between them. By decreasing the path-length, the absorbance due to the solvent will be reduced proportionately.
Your main problem is, that if you could you would measure change induced on protein by DMSO (denaturation) in first place.
I agree with Titus. You´ll need to make sure that you don´t just measure effect of DMSO on the protein.
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