Asynchronous HeLa cells were formaldehyde fixed, permeabilized with sodium citrate and Triton® X-100 (Dow Chemical Co) and blocked with PBS containing 2.5% BSA. (A) Cells were labeled with the anti-H3K27me3S28p crude serum (diluted 1:200 and incubated for 1 hour at room temperature) followed by labeled goat anti-rabbit secondary antibody (Fisher Scientific). (B) Nuclei were stained using the DNA-specific stain DAPI. Phosphorylation of H3 on serine 28 is increased during late G2 phase and reaches a maximum in metaphase cells. This may explain the different staining intensities of different cells.
|Tested species reactivity||Human|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Raised against histone H3 containing the trimethylated lysine 27 and the phosphorylated serine 28 (H3K27me3S28p), using a KLH conjugated synthetic peptide.|
|Storage buffer||whole serum|
|Contains||<0.1% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||1 ug|
|ELISA (ELISA)||1:100 to 1:500|
|Western Blot (WB)||1:250|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|ChIP assay (ChIP)||See 1 publications below|
Histone H3 is one of the DNA-binding proteins found in the chromatin of all eukaryotic cells. H3 along with four core histone proteins binds to DNA forming the structure of the nucleosome. Histones play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. Post tranlationally, histones are modified in a variety of ways to either directly change the chromatin structure or allow for the binding of specific transcription factors. The N-terminal tail of histone H3 protrudes from the globular nucleosome core and can undergo several different types of post-translational modification that influence cellular processes. These modifications include the covalent attachment of methyl or acetyl groups to lysine and arginine amino acids and the phosphorylation of serine or threonine.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Terminal epidermal differentiation is regulated by the interaction of Fra-2/AP-1 with Ezh2 and ERK1/2.
49-1015 was used in ChIP assay to assess the interaction between Ezh2 and ERK1/2 with Fra-2/AP-1 and regulation of terminal epidermal differentiation
|Wurm S,Zhang J,Guinea-Viniegra J,García F,Muñoz J,Bakiri L,Ezhkova E,Wagner EF||Genes and development (29:144)||2015|