Human HDAC1 protein, His tag

Histone deacetylase 1 (HDAC1) is an enzyme which belongs to the histone deacetylase/acuc/apha family and is a component of the histone deacetylase complex. Histone acetylation and deacetylation, catalyzed by multisubunit complexes, play a key role in the regulation of eukaryotic gene expression. HDAC1 also interacts with retinoblastoma tumor-suppressor protein and this complex is a key element in the control of cell proliferation and differentiation. Besides, Specificity Protein 1 (Sp1) has been identified as an interactor of HDAC1, thus a binding ELISA assay was conducted to detect the interaction of recombinant human HDAC1 and recombinant human Sp1. Briefly, HDAC1 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to Sp1-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-HDAC1 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 37ºC. Finally, add 50 μl stop solution to the wells and read at 450nm immediately. The binding activity of of HDAC1 and Sp1 was in a dose dependent manner.