Hi, i am looking to use Hsp90 Inhibitor Geldanamycin. I have optimized the assay where i have a substrate protein and this chaperone and i have monitored the prevention of aggregation using a Peltier based flourimeter. Now i have added GA(Geldanamycin) at increasing micromolar concentrations( 20um,40um,60um...) but this compound shows very high absorbance itself when checked, without any proteins. The scale of the aggregtion in the software is from 1 to 1000 A.U. and at 80um only GA gives absorbance around 800-850 A.U. which makes it impossible to monitor the effect it has on aggregation prevention. Does this compound absorb at 500nm or is it because of the DMSO(
It might not be absorbance you are observing but light-scattering. The GA may be starting to crash out. If this is the case, then changing the excitation/emission wavelength will not help. Lower the concentration of GA, increase DMSO or try more soluble version of GA.
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