Western blot analysis of NCoR2 was performed by loading 25 ug of Hela (lane 1) and Neuro-2a (lane 2) cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4°C overnight. The membrane was probed with a NCoR2 monoclonal antibody (Product # MA1-843) at a dilution of 1:100 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at ~220-250 kDa in Hela cells.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Synthetic peptide corresponding to residues C G G A(2016) S A S D P H R E K T Q(2027) of human SMRT.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||Assay Dependent|
|Immunohistochemistry (IHC)||Assay Dependent|
|Western Blot (WB)||4 µg/ml|
* 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.
|Immunocytochemistry (ICC)||See 1 publications below|
MA1-843 detects silencing mediator of retinoic acid and thyroid hormone receptor (SMRT) from human cells and tissues.
MA1-843 has been successfully used in Western blotting and immunohistochemical procedures. By Western blot, this antibody detects ~250 kDa and ~220 kDa proteins representing endogenous levels of both known isoforms of SMRT in HeLa cell extracts. This antibody does not react with mouse tissue in Western blot applications.
The MA1-843 immunogen is a synthetic peptide corresponding to residues C G G A(2016) S A S D P H R E K T Q(2027) of human SMRT. MA1-843 immunizing peptide (Cat. # PEP-043) is available for use in neutralization and control experiments.
Steroid and thyroid hormones and retinoic acid regulate a complex array of gene expression activity via intracellular receptor transcription factors belonging to the ligand-dependent nuclear receptor superfamily. Adding to the complexity of function of these transcription factors are associated proteins known as coactivators and corepressors which, as their names suggest, enhance or depress transcription activity of the nuclear receptor with which they associate. Silencing mediator of retinoic acid and thyroid hormone receptor (SMRT) and nuclear receptor corepressor (N-CoR) are related transcriptional corepressors which contain two distinct domains capable of interacting with unliganded nuclear receptors to repress their basal transcriptional activity.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Regulation of subnuclear localization is associated with a mechanism for nuclear receptor corepression by RIP140.
MA1-843 was used in immunocytochemistry to investigate the role of RIP140 during subnuclear localization regulation.
|Tazawa H,Osman W,Shoji Y,Treuter E,Gustafsson JA,Zilliacus J||Molecular and cellular biology (23:4187)||2003|