Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade (ab63766)

Overview

  • Product nameAnti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP GradeSee all TATA binding protein TBP primary antibodies ...
  • Description
    Rabbit polyclonal to TATA binding protein TBP - Nuclear Loading Control and ChIP Grade
  • Tested applicationsICC/IF, IP, ChIP, IHC-P, WB more details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chicken, Cow
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human TATA binding protein TBP.

    (Peptide available as ab64117.)

  • Positive control
    • This antibody gave a positive signal in the following lysates: HeLa Whole Cell, HepG2 Whole Cell, HeLa Nuclear (data not shown), HepG2 Nuclear (data not shown), Mouse Testis Tissue, NIH 3T3 Whole Cell, Rat Testis Tissue. This antibody gave a positive result in IHC in the following FFPE tissue: Human breast ductal carcinoma.

Properties

Applications

Our Abpromise guarantee covers the use of ab63766 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
ICC/IF 1/500.
IP Use at an assay dependent concentration.
ChIP Use 5 µg for 25 µg of chromatin.
IHC-P Use a concentration of 10 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 45 kDa (predicted molecular weight: 38 kDa).Can be blocked with Human TATA binding protein TBP peptide (ab64117).

Target

  • FunctionGeneral 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. 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. The rate of PIC formation probably is primarily dependent on the rate of association of SL1 with the rDNA promoter. SL1 is involved in stabilization of nucleolar transcription factor 1/UBTF on rDNA.
  • Tissue specificityWidely expressed, with levels highest in the testis and ovary.
  • Involvement in diseaseDefects in TBP are the cause of spinocerebellar ataxia type 17 (SCA17) [MIM:607136]. Spinocerebellar ataxia is a clinically and genetically heterogeneous group of cerebellar disorders. Patients show progressive incoordination of gait and often poor coordination of hands, speech and eye movements, due to degeneration of the cerebellum with variable involvement of the brainstem and spinal cord. SCA17 is an autosomal dominant cerebellar ataxia (ADCA) characterized by widespread cerebral and cerebellar atrophy, dementia and extrapyramidal signs. 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.
  • Sequence similaritiesBelongs to the TBP family.
  • Cellular localizationNucleus.
  • Target information above from: UniProt accession P20226 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
    • GTF2D antibody
    • GTF2D1 antibody
    • HDL4 antibody
    • MGC117320 antibody
    • MGC126054 antibody
    • MGC126055 antibody
    • SCA17 antibody
    • TATA binding factor antibody
    • TATA box factor antibody
    • TATA sequence binding protein antibody
    • TATA sequence-binding protein antibody
    • TATA-binding factor antibody
    • TATA-box binding protein N-terminal domain antibody
    • TATA-box factor antibody
    • TATA-box-binding protein antibody
    • TBP antibody
    • TBP_HUMAN antibody
    • TFIID antibody
    • Transcription initiation factor TFIID TBP subunit antibody
    see all

Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade images

  • All lanes : Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade (ab63766) at 1 µg/ml

    Lane 1 : Testis (Mouse) Tissue Lysate
    Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : Testis (Rat) Tissue Lysate
    Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Nuclear Lysate (ab14660)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 38 kDa
    Observed band size : 38,45 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 55 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 3 minutes
  • Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab63766 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach for active loci and Taqman approach for inactive loci). Primers and probes are located in the first kb of the transcribed region.
  • TATA binding protein TBP was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Rabbit polyclonal to TATA binding protein TBP and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, HepG2 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab63766.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 45kDa: TATA binding protein TBP.
  • All lanes : Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade (ab63766) at 1 µg/ml

    Lane 1 : Human TATA binding protein TBP full length protein (ab81897) at 0.1 µg
    Lane 2 : Human TATA binding protein TBP full length protein (ab81897) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 40 kDa


    Exposure time : 10 seconds
  • IHC image of TATA binding protein TBP staining in Human breast ductal carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab63766, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • ab63766 (1/500) staining TATA binding protein (TBP) in HeLa cells (green). Cells were fixed with paraformaldehyde, permeabilised with 0.5% Triton X100/PBS and counterstained with DAPI (red) in order to highlight the nucleus. Please refer to abreview for further experimental details.

    See Abreview

  • All lanes : Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade (ab63766) at 1 µg/ml

    Lane 1 : HeLa Whole Cell Lysate
    Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 38 kDa
    Observed band size : 45 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 60 kDa. We are unsure as to the identity of these extra bands.

References for Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade (ab63766)

This product has been referenced in:
  • Reddy GR  et al. GnRH increases c-Fos half-life contributing to higher FSHß induction. Mol Endocrinol 27:253-65 (2013). Read more (PubMed: 23275456) »
  • Fontenot E  et al. A novel monoclonal antibody to secreted frizzled-related protein 2 inhibits tumor growth. Mol Cancer Ther 12:685-95 (2013). WB ; Human . Read more (PubMed: 23604067) »

See all 5 Publications for this product

Product Wall

Thank you for contacting us.

Both nuclear loading controls you are referring are suitable to use with human lysates.

Please make sure the protein to be detected in the blot has a different molecular weight than the expected for TBP ...

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Thank you for your enquiry.

I am sorry to confirm that as far as we are aware,these three TPB antibodies(ab818, ab51841 and ab63766) have not been tested with samples from Methanococcoides burtonii. All tested and guaranteedspecies cross-rea...

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Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton X100 in PBS
Username

Dr. Kirk McManus

Verified customer

Submitted Feb 24 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"