Immunofluorescence analysis of NCoR2 was done on 70% confluent log phase PC-3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with NCoR2 (1212) Mouse Monoclonal Antibody (MA1842) at 2ug/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested 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 *|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|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.
MA1-842 detects silencing mediator of retinoic acid and thyroid hormone receptor (SMRT) from human cells. This antibody fails to detect rat SMRT.
MA1-842 has been successfully used in Western blot and immunohistochemical procedures. By Western blot, this antibody detects ~250 kDa and ~220 kDa proteins representing endogenous levels of both known isoforms of human SMRT in HeLa cell extracts. Immunohistochemical staining of SMRT in HeLa cells with MA1-842 results in nuclear staining.
The MA1-842 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. MA1-842 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.