HDT1 | Histone deacetylase

Recombinant part of Arabidopsis thaliana HDT1 Q9FVE6, At3g44750

application example

50 µg of Arabidopsis T87 nuclear proteins, extracted with Nuclei Lysis Buffer (1% SDS, 50mM Tris-HCl, 10mM EDTA, Complete EDTA-free) or 50 µg of total Arabidopsis T87 proteins, extracted with Laemmli Buffer from grinded in liquid nitrogen material, were separated on 12 % SDS-PAGE and blotted 1,5h to PVDF membrane (WestranS 0,20 µm, Whatman). Blots were blocked with 5% fat free milk in TBST for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 in 2,5% fat-free milk in TBST overnight at 40°C with agitation. The antibody solution was decanted and the blot was washed with TBST three times for 10 min. and blocked in 10% fat-free milk in TBST for 10 min. Next, blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:5 000 in 5% fat-free milk in TBST for 2h at RT with agitation. The blot was washed six times for 10 min. with TBST and developed for 5 min with ECL+ (Amersham) according to the manufacturers instructions. Exposure time was 1 min.

Courtesy of Msc. Daniel Buszewicz, PAN, Poland

HDT1 (HD2A) is probably mediating in the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4). Histone deacetylation gives a tag for epigenetic repression and plays an important role in cell cycle progression, transcriptional regulation and developmental events. This protein is also involved in rRNA gene silencing in nucleolar dominance. Synonymes:HD-tuins protein 1, Histone deacetylase 2a.