Mouse differentiated ES cells were formaldehyde fixed, permeabilized with Triton® X-100 (Dow Chemical Co) and then blocked with PBS containing BSA. Cells were labeled with the anti-HDAC1 (diluted 1:200 and incubated for 45 minutes at room temperature) followed by labeled goat anti-rabbit secondary antibody. Subsequently, RNA FISH (fluorescence in situ hybridization) was performed to detect Xist RNA (green signal). Nuclei were DAPI-stained to specifically label the DNA. As expected, both DAPI and HDAC1 staining occur in the nucleus of the cells. Note the broadly dispersed characteristic pattern of this protein mark in interphase chromatin but its exclusion from constitutive heterochromatin in mouse cells.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||raised in rabbits against the C-terminal region of human HDAC1, using a KLH-conjugated synthetic peptide.|
|Contains||<0.1% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||2.4 ug|
|ELISA (ELISA)||1:100 to 1:300|
* 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.
|Western Blot (WB)||See 1 publications below|
Chromatin is a highly specialized structure composed of tightly compacted chromosomal DNA. Gene expression within the nucleus is controlled, in part, by a host of protein complexes which continuously pack and unpack the chromosomal DNA. One of the known mechanisms of this packing and unpacking process involves the acetylation and deacetylation of the histone proteins comprising the nucleosomal core. Acetylated histone proteins confer accessibility of the DNA template to the transcriptional machinery for expression. Histone deacetylases (HDACs) are chromatin remodeling factors that deacetylate histone proteins and thus, may act as transcriptional repressors. HDACs are classified by their sequence homology to the yeast HDACs and there are currently 2 classes. Class I proteins are related to Rpd3 and members of class II resemble Hda1p. HDAC1 is a class I histone deacetylase containing 482 amino acid residues. HDAC1 has been shown to interact directly with transcription factors and has been shown to deacetylate histone proteins H3 and H4.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Transcriptional repression of ER through hMAPK dependent histone deacetylation by class I HDACs.
49-1025 was used in western blot to study the relationship between hyperactivation of MAPK and the estrogen receptor in breast cancer cells
|Plotkin A,Volmar CH,Wahlestedt C,Ayad N,El-Ashry D||Breast cancer research and treatment (147:249)||2014|