HTA9 | Probable histone H2A variant 3
AS10 718 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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14.2 kDa (Arabidopsis thaliana)
1) hta9, hta11 double mutant, about 1 ug protein; 2) mutant hta9, hta11 about 2 ug protein; 3) wild-type about 2-3 ug protein; 4) wild-type about 4-5 ug protein from Arabidopsis thaliana.
Protein from Arabidopsis histone preparation (acid extracted, precipitated with acetone and resuspended in urea). Denatured with SDS-PAGE buffer at 90C for 2 min, separated on 15% SDS-PAGE and blotted 1h to PVDF tank transfer. Blots were blocked with PBS + 3% BSA overnight at 4C with agitation. Blot was incubated in the primary antibody at a dilution of 1:1 000 in PBS + 3% BSA for 1h at RT with agitation. The antibody solution was decanted and the blot was rinsed once with PBS-T for 5 min. Blot was incubated in secondary antibody PBS + 3% BSA (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera, AS09 602) diluted to 1:25 000 for 1h at RT with agitation. The blot was washed 3-4 times with PBS-T for 5 min. The blot was developed with Immobilion (Millipore) and exposed on an X-ray film for 1 min.
Variant histone H2A may replace conventional H2A in a subset of nucleosomes. Synonymes: H2A.F/Z 3
Gómez-Zambrano et al. (2019). The repressive role of Arabidopsis H2A.Z in transcriptional regulation depends on AtBMI1 activity. Nat Commun. 2019 Jun 27;10(1):2828. doi: 10.1038/s41467-019-10773-1.
Gómez-Zambrano et al. (2018). Arabidopsis SWC4 Binds DNA and Recruits the SWR1 Complex to Modulate Histone H2A.Z Deposition at Key Regulatory Genes. Mol Plant. 2018 Mar 29. pii: S1674-2052(18)30122-9. doi: 10.1016/j.molp.2018.03.014.