Active Histone Deacetylase 6 (HDAC6)

Histone Deacetylase 6 (HDAC6) belongs to class II of the histone deacetylase/acuc/apha family. It contains an internal duplication of two catalytic domains that appear to function independently of each other. This protein possesses histone deacetylase activity and represses transcription. HDAC6 also affects transcription and translation by regulating the heat-shock protein 90 (Hsp90) and stress granules (SGs), respectively. Besides, Sirtuin 2 (SIRT2) has been identified as an interactor of HDAC6, thus a binding ELISA assay was conducted to detect the interaction of recombinant human HDAC6 and recombinant human SIRT2. Briefly, HDAC6 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μl were then transferred to SIRT2-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-HDAC6 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of HDAC6 and SIRT2 was shown in Figure 1, and this effect was in a dose dependent manner.