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17-10057

Sigma-Aldrich

ChIPAb+ SMRT - ChIP Validated Antibody and Primer Set

ascites fluid, from mouse

Synonyme(s) :

CTG repeat protein 26, Silencing mediator of retinoic acid and thyroid hormone receptor, T3 receptor-associating factor, Thyroid-, retinoic-acid-receptor-associated corepressor, nuclear receptor co-repressor 2, silencing mediator for retinoid and thyroid

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About This Item

Code UNSPSC :
12352203
eCl@ss :
32160702
Nomenclature NACRES :
NA.32

Source biologique

mouse

Niveau de qualité

Forme d'anticorps

ascites fluid

Clone

monoclonal

Espèces réactives

human, mouse, rat

Fabricant/nom de marque

ChIPAb+
Upstate®

Technique(s)

ChIP: suitable
electrophoretic mobility shift assay: suitable
immunocytochemistry: suitable
immunohistochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable

Isotype

IgG2aκ

Numéro d'accès NCBI

Numéro d'accès UniProt

Conditions d'expédition

dry ice

Informations sur le gène

Description générale

All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
The ChIPAb+ SMRT set includes the SMRT antibody, a negative control mouse ascites, and qPCR primers which amplify a 299 bp region of human IκBα promoter. The SMRT and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of SMRT-associated chromatin.
The Silencing mediator of retinoic acid & thyroid hormone receptor protein, commonly called the SMRT protein mediates the transcriptional repression of some nuclear receptors by promoting chromatin condensation, thus preventing access of the basal transcription machinery. Consistent with this activity, this protein is known to form a large corepressor complex containing SIN3A/B and histone deacetylases HDAC1 and HDAC2. SMRT is also a component of the N-Cor repressor (nuclear receptor corepressor), a multi-subunit complex minimally composed of NCOR1, NCOR2, HDAC3, TBL1X, TBL1R, CORO2A and GPS2. 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.

Spécificité

Recognizes SMRT.

Immunogène

Recombinant protein corresponding to human SMRT.

Application

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from serum starved, TNFα treated (20 ng/mL, 30 min) HeLa cells (~3 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µL of Negative Ascites or 2 µL Anti-SMRT) and the Magna ChIP G Kit (Cat. # 17-611).
Successful immunoprecipitation of SMRT associated DNA fragments was verified by qPCR using ChIP Primers, IĸBα promoter as a positive locus, and β-Actin promoter primers as a negative locus (Please see figures).
Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
Please refer to the EZ-Magna ChIP A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.

Western Blot Analysis:
Representative lot data.
Human brain lysate was resolved by electrophoresis, transferred to PVDF membranes and probed with Anti-SMRT (1:1000 dilution).
Proteins were visualized using a Goat Anti-Mouse IgG conjugated to HRP and a chemiluminescence detection system (Please see figures).
Research Category
Epigenetics & Nuclear Function
Research Sub Category
RNA Metabolism & Binding Proteins
The ChIPAb+ SMRT set includes the SMRT antibody, a negative control mouse ascites & qPCR primers which amplify a 299 bp region of human IκBα promoter.

Conditionnement

25 assays per set. Recommended use: ~2 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).

Qualité

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from serum starved, TNFα treated (20 ng/mL, 30 min) HeLa cells (~3 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µL of Negative Ascites or 2 µL Anti-SMRT) and the Magna ChIP® G Kit (Cat. # 17-611).
Successful immunoprecipitation of SMRT associated DNA fragments was verified by qPCR using ChIP Primers, IĸBα promoter (Please see figures).

Please refer to the EZ-Magna ChIP A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.

Description de la cible

~ 275 kDa

Forme physique

Anti-SMRT (mouse monoclonal). One vial containing 50 µL of unpurified mouse monoclonal ascites with 0.05% sodium azide. Store at -20°C.

Negative Ascites (mouse). One vial containing 50 µL of mouse ascites with 0.05% sodium azide. Store at -20°C.

ChIP Primers, IκBα promoter. One vial containing 75 μL of 5 μM of each primer specific for human IĸBα promoter. Store at -20°C.
FOR: GAC GAC CCC AAT TCA AAT CG
REV: TCA GGC TCG GGG AAT TTC C
Unpurified

Stockage et stabilité

Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Remarque sur l'analyse

Control
Includes negative control mouse ascites and primers specific for human IκBα promoter.

Informations légales

MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Clause de non-responsabilité

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Code de la classe de stockage

10 - Combustible liquids


Certificats d'analyse (COA)

Recherchez un Certificats d'analyse (COA) en saisissant le numéro de lot du produit. Les numéros de lot figurent sur l'étiquette du produit après les mots "Lot" ou "Batch".

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Retrouvez la documentation relative aux produits que vous avez récemment achetés dans la Bibliothèque de documents.

Consulter la Bibliothèque de documents

Jonathan H Kahn et al.
Endocrinology, 161(10) (2020-08-10)
The Silencing Mediator of Retinoid and Thyroid Hormone Receptors (SMRT) is a nuclear corepressor, regulating the transcriptional activity of many transcription factors critical for metabolic processes. While the importance of the role of SMRT in the adipocyte has been well-established
Margo P Emont et al.
Molecular and cellular endocrinology, 407, 52-56 (2015-03-15)
Local modulation of glucocorticoid action in adipocytes regulates adiposity and systemic insulin sensitivity. However, the specific cofactors that mediate glucocorticoid receptor (GR) action in adipocytes remain unclear. Here we show that the silencing mediator of retinoid and thyroid hormone receptors
Lauren B Auriemma et al.
Epigenetics & chromatin, 5(1), 2-2 (2012-01-14)
The tumor suppressor menin (MEN1) is mutated in the inherited disease multiple endocrine neoplasia type I, and has several documented cellular roles, including the activation and repression of transcription effected by several transcription factors. As an activator, MEN1 is a
Pragya Sharma et al.
The Journal of biological chemistry, 288(23), 16321-16333 (2013-05-01)
Secretory phospholipase A2 group IIa (PLA2g2a) is associated with inflammation, hyperlipidemia, and atherogenesis. Transcription of the PLA2g2a gene is induced by multiple cytokines. Here, we report the surprising observation that thyroid hormone (T3) inhibited PLA2g2a gene expression in human and

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