i want to study the effect of bsa and hsa protein on my molecule's property vie uv spectra,by changing protein's concentration.how much stock solution do i need to prepare. do i prepare in water or buffer solutioin .
You should take into account that both the compound and the protein have UV spectra. If the spectra overlap, as is often the case, you will have to subtract the background spectrum from one component to see the spectrum of the other. If possible, dissolve both the compound and the protein in the same buffer in order to control the pH and to avoid having to correct for effect of the solvent on the spectra.
thank you very much but my compund is not soluble in buffer so i have to go threough polar organic solvent like methanol,if i choolse methanol than protein will be denatured so can i carry out my experiment with denatured protein sol or can you suggest me any paper where max solubility of protein in methanol is reported so than i can cieted that
Do not use methanol. Use DMSO (dimethyl sulfoxide). BSA and HSA should be able to tolerate at least a few % DMSO by volume. Keep the DMSO concentration constant. Make sure to include the same amount of DMSO in blanks.
i cant use DMSO becouse of % of DMSO/water is very high for making solution and also i dont want to make it tri-solvent system since i have to use methanol. can you sugget me any limiting concentration or weight of BSA or HSA to make solution in methanolic solution.i have find some paper where they make 0.15 uM solution in methanol and it is ok.
i can only use methanol/water or methanol/glycerol.... what do you suggest.here i attach file support the methanolic system to 0.15 uM solution
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