HTA9 | Probable histone H2A variant 3

AS10 718 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

Product Information

Immunogen

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana HTA9 UniProt: Q9C944, TAIR: At1g52740

Host Rabbit

Clonality Polyclonal

Purity Affinity purified serum in PBS, pH 7.4

Format Lyophilized

Quantity 50 µg

Reconstitution For reconstitution add 50 µl of sterile water.

Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

Tested applications Chromatin Immunoprecipitation (ChIP), Immunoprecipitation (IP), Western blot (WB)

Recommended dilution 1: 100 (IP), 1 : 1000 (WB)

Expected | apparent MW

14.2 kDa (Arabidopsis thaliana)

Reactivity

Confirmed reactivity Arabidopsis thaliana

Not reactive in Nicotiana tabacum

Application examples

Application examples Application example

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.

Additional information

Related products

collection of antibodies to DNA/RNA/cell cycle

Plant protein extraction buffer

Secondary antibodies

Background

Variant histone H2A may replace conventional H2A in a subset of nucleosomes. Synonymes: H2A.F/Z 3

Product citations

Selected references Kralemann et al. (2020). Removal of H2Aub1 by ubiquitin-specific proteases 12 and 13 is required for stable Polycomb-mediated gene repression in Arabidopsis. Genome Biol. 2020 Jun 16;21(1):144.doi: 10.1186/s13059-020-02062-8.
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.