In ELISA analysis ab4633 recognises di methyl histone H2A K95 but not the corresponding unmodified or histone H4 di methyl K79 peptides. However, ab4633 does to an extent also recognise a di methyl K99 peptide in ELISA analysis.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml.
1/1000. Detects a band of approximately 15 kDa.
Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.
Belongs to the histone H2A family.
The chromatin-associated form is phosphorylated on Thr-121 during mitosis. Deiminated on Arg-4 in granulocytes upon calcium entry. Monoubiquitination of Lys-120 by RING1 and RNF2/RING2 complex gives a specific tag for epigenetic transcriptional repression and participates in X chromosome inactivation of female mammals. It is involved in the initiation of both imprinted and random X inactivation. Ubiquitinated H2A is enriched in inactive X chromosome chromatin. Ubiquitination of H2A functions downstream of methylation of 'Lys-27' of histone H3. Monoubiquitination of Lys-120 by RNF2/RING2 can also be induced by ultraviolet and may be involved in DNA repair. Following DNA double-strand breaks (DSBs), it is ubiquitinated through 'Lys-63' linkage of ubiquitin moieties by the E2 ligase UBE2N and the E3 ligases RNF8 and RNF168, leading to the recruitment of repair proteins to sites of DNA damage. Monoubiquitination and ionizing radiation-induced 'Lys-63'-linked ubiquitination are distinct events. Phosphorylation on Ser-2 is enhanced during mitosis. Phosphorylation on Ser-2 by RPS6KA5/MSK1 directly represses transcription. Acetylation of H3 inhibits Ser-2 phosphorylation by RPS6KA5/MSK1. Symmetric dimethylation on Arg-4 by the PRDM1/PRMT5 complex may play a crucial role in the germ-cell lineage.
ICC/IF image of ab4633 stained human HEK 293 cells. The cells were methanol fixed (5 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab4633, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Western blot - Anti-Histone H2A (di methyl K95) antibody (ab4633)
Lane 1 : 1ug of calf thymus Histone prep Lane 2 : 1ug of calf thymus Histone prep Lane 3 : 1ug of calf thymus Histone prep with Human Histone H2A (di methyl K95) peptide (ab10127) at 2 µg Lane 4 : 1ug of calf thymus Histone prep with Human Histone H2A (di methyl K95) peptide (ab10127) at 2 µg
Secondary Lanes 1 - 3 : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 2000 Lane 4 : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/2000 dilution