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Merck
CN

17-10057

ChIPAb+ SMRT - ChIP Validated Antibody and Primer Set

ascites fluid, from mouse

别名:

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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关于此项目

UNSPSC代码:
12352203
eCl@ss:
32160702
NACRES:
NA.32

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生物来源

mouse

质量水平

100

抗体形式

ascites fluid

克隆

monoclonal

种属反应性

human, mouse, rat

制造商/商品名称

ChIPAb+
Upstate®

技术

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

同位素/亚型

IgG2aκ

NCBI登记号

NP_001070729.1

UniProt登记号

Q9Y618

运输

dry ice

基因信息

human ... NCOR2(9612), TRAC(28755)

一般描述

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.
~ 275 kDa

免疫原

Recombinant protein corresponding to human SMRT.

应用

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.

生化/生理作用

Recognizes SMRT.

包装

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

外形

Unpurified
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

制备说明

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.

分析说明

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

法律信息

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

免责声明

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.

储存分类代码

10 - Combustible liquids

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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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