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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| H, M, R | ICC, IHC, IP, WB, EMSA, ChIP |
| Description | |
|---|---|
| Catalogue Number | 17-10057 |
| Brand Family | Upstate |
| Trade Name |
|
| Description | ChIPAb+ SMRT - ChIP Validated Antibody and Primer Set |
| Overview | 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. |
| Alternate Names |
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| Background Information | 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. |
| Product Information | |
|---|---|
| Format | Ascites |
| Control |
|
| Presentation | 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 |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | 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. |
| Key Applications |
|
| Application Notes | 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). |
| Biological Information | |
|---|---|
| Immunogen | Recombinant protein corresponding to human SMRT. |
| Host | Mouse |
| Specificity | Recognizes SMRT. |
| Isotype | IgG2aκ |
| Species Reactivity |
|
| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | The protein encoded by this gene interacts with certain retinoid and thyroid hormone receptors involved in transcriptional repression. The encoded protein is part of a large protein complex that facilitates chromatin condensation and repression. Two transcript variants encoding different isoforms have been found for this gene. |
| Gene Symbol |
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| Purification Method | Unpurified |
| UniProt Number |
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| UniProt Summary | FUNCTION: Mediates the transcriptional repression activity of some nuclear receptors by promoting chromatin condensation, thus preventing access of the basal transcription. SIZE: 2517 amino acids; 274034 Da SUBUNIT: Interacts with HDAC7 (By similarity). Forms a large corepressor complex that contains SIN3A/B and histone deacetylases HDAC1 and HDAC2. This complex associates with the thyroid (TR) and the retinoid acid receptors (RAR) in the absence of ligand, and may stabilize their interaction with TFIIB. The SRMT isoform interacts with HDAC10. Interacts with MINT. Component of the N-Cor repressor complex, at least composed of NCOR1, NCOR2, HDAC3, TBL1X, TBL1R, CORO2A and GPS2. Interacts with CBFA2T3. SUBCELLULAR LOCATION: Nucleus. TISSUE SPECIFICITY: Ubiquitous. High levels of expression are detected in lung, spleen and brain. DOMAIN: SwissProt: Q9Y618 The N-terminal region contains repression functions that are divided into three independent repression domains (RD1, RD2 and RD3). The C-terminal region contains the nuclear receptor- interacting domains that are divided in two separate interaction domains (ID1 and ID2). & The two interaction domains (ID) contain a conserved sequence referred to as the CORNR box. This motif is required and sufficient to permit binding to unligated TR and RARS. Sequences flanking the CORNR box determine nuclear hormone receptor specificity. SIMILARITY: Belongs to the N-CoR nuclear receptor corepressors family. & Contains 2 SANT domains. |
| Molecular Weight | ~ 275 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | 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. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | 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. |
| Packaging Information | |
|---|---|
| Material Size | 25 assays |
| Material Package | 25 assays per set. Recommended use: ~2 μL of antibody per chromatin immunoprecipitation (dependent upon biological context). |