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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| Ch, H, M, Xn, F | WB, ChIP |
| Description | |
|---|---|
| Catalogue Number | 17-10098 |
| Brand Family | Upstate |
| Trade Name |
|
| Description | ChIPAb+ TATA Binding Protein (TBP) - 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+ TATA Binding Protein (TBP) set includes the TATA Binding Protein (TBP) antibody, a negative control mouse ascites, and qPCR primers which amplify a 166 bp region of human GAPDH promoter. The TATA Binding Protein (TBP) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of TATA Binding Protein (TBP)-associated chromatin. |
| Alternate Names |
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| Background Information | TATA-box Binding Protein (TBP) is a general transcription factor that functions at the core of the DNA-binding multiprotein factor TFIID. Binding of TFIID to the TATA box is the initial transcriptional step of the pre-initiation complex (PIC). TBP plays a role in the activation of eukaryotic genes transcribed by RNA polymerase II and is a component of the transcription factor SL1/TIF-IB complex, which is involved in the assembly of the PIC (preinitiation complex) during RNA polymerase I-dependent transcription (Unitprot). |
| Product Information | |
|---|---|
| Format | Ascites |
| Control |
|
| Presentation | Anti-TATA Binding Protein (TBP) (mouse monoclonal). One vial containing 50 μL ascites containing 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, GAPDH promoter. One vial containing 75 μL of 5 μM each primer specific for human GAPDH. Store at -20°C. FOR: TAC TAG CGG TTT TAC GGG CG REV: TCG AAC AGG AGG AGC AGA GAG CGA |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | The ChIPAb+ TATA Binding Protein (TBP) set includes the TATA Binding Protein (TBP) antibody, a negative control mouse ascites & qPCR primers which amplify a 166 bp region of human GAPDH promoter. |
| Key Applications |
|
| Application Notes | Chromatin Immunoprecipitation: Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µL of either Negative Ascites or 2 µL Anti-TATA Binding Protein (TBP) and the Magna ChIP™ G Kit (Cat. # 17-611). Successful immunoprecipitation of TBP associated DNA fragments was verified by qPCR using ChIP Primers, human GAPDH promoter as a positive locus, and GAPDH coding 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™ G (Cat. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details. Western Blot Analysis: Representative lot data. HeLa lysates were resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-TATA Binding Protein (TBP) (1:1000 dilution). Proteins were visualized using a secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures). Immunohistochemistry: A 1:1,000-1:5,000 dilution of a previous lot was used in immunohistochemistry. Immunocytochemistry: A 1:1,000-1:5,000 dilution of a previous lot was used in immunocytochemistry. |
| Biological Information | |
|---|---|
| Immunogen | Full length recombinant human TBP. |
| Host | Mouse |
| Specificity | Recognizes TATA Binding Protein (TBP). |
| Isotype | IgG1 |
| Species Reactivity |
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| Species Reactivity Note | Human, mouse, chicken, xenopus and zebrafish. Does not react with Drososphila. Reactivity with other species has not been confirmed. |
| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | Initiation of transcription by RNA polymerase II requires the activities of more than 70 polypeptides. The protein that coordinates these activities is transcription factor IID (TFIID), which binds to the core promoter to position the polymerase properly, serves as the scaffold for assembly of the remainder of the transcription complex, and acts as a channel for regulatory signals. TFIID is composed of the TATA-binding protein (TBP) and a group of evolutionarily conserved proteins known as TBP-associated factors or TAFs. TAFs may participate in basal transcription, serve as coactivators, function in promoter recognition or modify general transcription factors (GTFs) to facilitate complex assembly and transcription initiation. This gene encodes TBP, the TATA-binding protein. A distinctive feature of TBP is a long string of glutamines in the N-terminal. This region of the protein modulates the DNA binding activity of the C terminus, and modulation of DNA binding affects the rate of transcription complex formation and initiation of transcription. Mutations that expand the number of CAG repeats encoding this polyglutamine tract, and thus increase the length of the polyglutamine string, are associated with spinocerebellar ataxia 17, a neurodegenerative disorder classified as a polyglutamine disease. |
| Gene Symbol |
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| Non-Reactive Species |
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| Purification Method | Unpurified |
| UniProt Number |
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| UniProt Summary | FUNCTION: SwissProt: P20226 # General transcription factor that functions at the core of the DNA-binding multiprotein factor TFIID. Binding of TFIID to the TATA box is the initial transcriptional step of the pre- initiation complex (PIC), playing a role in the activation of eukaryotic genes transcribed by RNA polymerase II. SIZE: 339 amino acids; 37698 Da SUBUNIT: Belongs to the TFIID complex together with the TBP- associated factors (TAFs). Component of the transcription factor SL1/TIFIB complex, composed of TBP and at least TAF1A, TAF1B TAF1C, and TAF3. Binds DNA as monomer. Interacts with TAFs, TFIIB, NCOA6, DRAP1, DR1 and ELF3. Interacts with SPIB, SNAPC1, SNAPC2 and SNAPC4. Interacts with HIV-1 Tat. Interacts with UTF1 which acts as a coactivator of ATF2 transcriptional activity. SUBCELLULAR LOCATION: Nucleus. DISEASE: SwissProt: P20226 # Defects in TBP are the cause of spinocerebellar ataxia 17 (SCA17) [MIM:607136]. Spinocerebellar ataxia is a clinically and genetically heterogeneous group of autosomal dominant cerebellar ataxias (ADCA). Patients show progressive incoordination of gait and often poor coordination of hands, speech and eye movements. Spinocerebellar ataxia is caused by degeneration of the cerebellum with variable involvement of the brainstem and spinal cord. The molecular defect in SCA17 is the expansion of a CAG repeat in the coding region of TBP. Longer expansions result in earlier onset and more severe clinical manifestations of the disease. SIMILARITY: SwissProt: P20226 ## Belongs to the TBP family. |
| Molecular Weight | ~42 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Chromatin Immunoprecipitation: Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µL of either Negative Ascites or 2 µL Anti-TATA Binding Protein (TBP) and the Magna ChIP™ G Kit (Cat. # 17-611). Successful immunoprecipitation of TATA Binding Protein (TBP)-associated DNA fragments was verified by qPCR using ChIP Primers, human GAPDH promoter (Please see figures). Please refer to the EZ-Magna ChIP™ G (Cat. # 17-409) 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). |