A Nature research paper reports how clusters of boron atoms can deliver small molecules into mammalian cells owing to their superchaotropic nature – meaning they are particularly good at disrupting hydrogen bonds between water molecules
= 585 nm) and concentrations were calculated by using a calibration curve with serial dilutions of TAMRA-R. For the complementary HPLC analysis, phosphate buffer was replaced by TBS and digitonin extraction was performed as indicated above. An aliquot of these extracts was used for β-hexosaminidase activity determination. Cytosolic extracts were lyophilized and resuspended in 1:10 volumes of HCN 100:0 ; 100:0→5:95 ) by monitoring the 555-nm absorbance of the TAMRA chromophore.
The quality of fractionation was assessed by lysosomal β-hexosaminidase activity, using 4-nitrophenyl 2-acetamido-2-deoxy-β--glucopyranoside as substrate. Briefly, 20 µl of extract was incubated with 80 µl of 7.5 mM substrate in 100 mM citrate buffer, pH 4.7, for 40 min at 37 °C, and the reaction was stopped by addition of 200 µl of 0.2 M Tris solution. Absorbance at 405 nm was measured in a plate reader. As blank, wells containing only the substrate were used.
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