The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 70 kDa (predicted molecular weight: 70 kDa).
Use a concentration of 5 µg/ml.
Use at an assay dependent concentration.
Use a concentration of 5 µg/ml.
FunctionEssential component of the BHC complex, a corepressor complex that represses transcription of neuron-specific genes in non-neuronal cells. The BHC complex is recruited at RE1/NRSE sites by REST and acts by deacetylating and demethylating specific sites on histones, thereby acting as a chromatin modifier. In the BHC complex, it serves as a molecular beacon for the recruitment of molecular machinery, including MeCP2 and SUV39H1, that imposes silencing across a chromosomal interval. Plays a central role in demethylation of Lys-4 of histone H3 by promoting demethylase activity of KDM1A on core histones and nucleosomal substrates. It also protects KDM1A from the proteasome.
Tissue specificityUbiquitously expressed.
Sequence similaritiesBelongs to the CoREST family. Contains 1 ELM2 domain. Contains 2 SANT domains.
DomainThe SANT domains may bridge the nucleosomal substrates and the demethylase KDM1A.
Post-translational modificationsPhosphorylated by HSV-1 protein kinases in case of infection.
Cellular localizationNucleus. Upon infection by HSV-1, it is partially translocated into the cytoplasm in an HSV-1-dependent manner.
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 4 : Jurkat (Human T cell lymphoblast-like cell line) Nuclear Lysate
Lysates/proteins at 20 µg per lane.
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size : 70 kDa Observed band size : 70 kDa Additional bands at : 15 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 10 minutes
ChIP - CoREST antibody (ab32631)
PCR was performed on a SCNA1 promoter following CoRest chromatin immunoprecipition using ab32631 at a concentration of 3ug/ml from 293T cells. Lane 1 - ab24166 Lane 2 - Beads only control Lane 3 - IgG only control Lane 4 - Input
CoREST was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to CoREST and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab32631. Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697). Band: 65kDa: CoREST.
IHC image of CoREST staining in Human normal tonsil FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab32631, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX
ICC/IF image of ab32631 stained MCF7 cells. The cells were 10% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32631, 5µg/ml) overnight at +4°C. 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) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, Hek293 and HepG2 cells at 5µg/ml.
References for Anti-CoREST antibody (ab32631)
This product has been referenced in:
Kim JY et al. Specificity in circadian clock feedback from targeted reconstitution of the NuRD corepressor. Mol Cell56:738-48 (2014).
Read more (PubMed: 25453762) »
Nicholson TB et al. A hypomorphic lsd1 allele results in heart development defects in mice. PLoS One8:e60913 (2013).
Read more (PubMed: 23637775) »