Tetramethylrhodamine-5-iodoacetamide (0.5, 1.0, 2.0, 3.0, 4.0, 5.0, and 7.5 mM dye) reveal different results for labeling carbonic anhydrase and myoglobin. At lower dye concentrations (up to 40:1 dye:thiol ratio) fluorescence labeling of only the carbonic anhydrase band is observed. However,as concentration increases, fluorescence labeling of both carbonic anhydrase and the lower myoglobin band is observed. Which indicating nonspecific labeling at these dye:thiol ratios[1].