Anti-TATA binding protein TBP antibody [1TBP18] - ChIP Grade (ab818)

Overview

  • Product nameAnti-TATA binding protein TBP antibody [1TBP18] - ChIP Grade
    See all TATA binding protein TBP primary antibodies
  • Description
    Mouse monoclonal [1TBP18] to TATA binding protein TBP - ChIP Grade
  • Tested applicationsChIP/Chip, ELISA, IHC-P, EMSA, IP, WB, Flow Cyt, ChIP, ICC/IF, ICC, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human

    Does not react with

    Saccharomyces cerevisiae, Xenopus laevis, Fruit fly (Drosophila melanogaster), Silk worm
  • Immunogen

    Synthetic peptide (Human).

  • EpitopeWithin amino acid residues 1-20 of human, mouse and rat TBP. The specific epitope for this antibody is accessible when the C-terminal domain of TBP is removed or when TBP is complexed with DNA.
  • Positive control
    • HeLa nuclear extract

Properties

Applications

Our Abpromise guarantee covers the use of ab818 in the following tested applications.

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

Application Abreviews Notes
ChIP/Chip Use at an assay dependent dilution.
ELISA 1/1000.
IHC-P Use at an assay dependent dilution.
EMSA Use at an assay dependent dilution.
IP Use at an assay dependent dilution.
WB 1/2000. Detects a band of approximately 38 kDa (predicted molecular weight: 38 kDa).
Flow Cyt Use 1µg for 106 cells.
ChIP Use 5-10 µg for 25 µg of chromatin.
ICC/IF Use at an assay dependent dilution.
ICC Use at an assay dependent dilution.
IHC-Fr Use at an assay dependent dilution.

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.
  • Information by UniProt
  • Database links
  • Alternative names
    • GTF2D antibody
    • GTF2D1 antibody
    • MGC117320 antibody
    • MGC126054 antibody
    • MGC126055 antibody
    • SCA 17 antibody
    • SCA17 antibody
    • TATA binding factor antibody
    • TATA box binding protein antibody
    • TATA box factor antibody
    • TATA sequence binding protein antibody
    • TATA sequence-binding protein antibody
    • TATA-binding factor antibody
    • TATA-box factor antibody
    • TATA-box-binding protein antibody
    • TBP antibody
    • TBP_HUMAN antibody
    • TF2D antibody
    • TFIID antibody
    • Transcription initiation factor TFIID TBP subunit antibody
    see all

Anti-TATA binding protein TBP antibody [1TBP18] - ChIP Grade images

  • Anti-TATA binding protein TBP antibody [1TBP18] - ChIP Grade (ab818) at 1/2000 dilution + Ros C cells with endogenous TBP

    Performed under reducing conditions.

    Predicted band size : 38 kDa
  • Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The ChIP was performed with 25 µg of chromatin, 8 µg of  ab818 (blue), and 20 µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The last wash was performed with final wash buffer containing 250 mM NaCl. The immunoprecipitated DNA was quantified by real time PCR (Taqman and sybr green approach). Primers and probes are located in the core promoter region of the genes.

  • ab818 staining TATA binding protein TBP by ELISA. The sample was purified from human AGS gastric carcinoma cell line and blocked with 5% BSA for 1 hour at 25°C. The primary antibody was used at 1/1000 dilution, and incubated with sample for 16 hour at 4°C. ab6729 AP conjugated rabbit polyclonal to mouse IgG was used as secondary, diluted at 1/1000.

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  • All lanes : Anti-TATA binding protein TBP antibody [1TBP18] - ChIP Grade (ab818) at 1/2000 dilution

    Lane 1 : Human Huh7 nuclear cell lysate
    Lane 2 : Human Huh7 cytoplast cell lysate
    Lane 3 : Human HepG2 nuclear cell lysate
    Lane 4 : Human HepG2 cytoplast cell lysate

    Lysates/proteins at 40 µg per lane.

    Secondary
    HRP-conjugated goat polyclonal to mouse IgG at 1/10000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 38 kDa
    Observed band size : 38 kDa


    Exposure time : 30 seconds

    This image is a courtesy of Chien Hsin Lee

    See Abreview

  • ab818 staining TATA binding protein TBP in Human stomach tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). The tissue underwent formaldehyde fixation before heat mediated antigen retrieval in 10mM Citrate buffer pH 6.0. Blocking was done in 5% serum for 1 hour at 23°C. The primary antibody was diluted 1/100 and incubated with the sample for 1 hour at 23°C. An HRP-conjugated Goat polyclonal to Mouse IgG was used undiluted as the secondary antibody..

     

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  • ab818 staining TATA binding protein TBP in NIH3T3 mouse fibroblast cells by Immunocytochemistry/ Immunofluorescence. The cells were fixed in formaldehyde, permeabilised in 0.025% Triton X and then blocked using 5% serum for 1 hour at 23°C. Samples were then incubated with primary antibody at 1/200 for 16 hours at 4°C. The secondary antibody used was a goat anti-mouse IgG conjugated to Alexa Fluor® 350 (blue) used at a 1/1000 dilution.

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  • Overlay histogram showing HeLA cells stained with ab818 (red line). The cells were fixed with 100% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab818, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was Mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References for Anti-TATA binding protein TBP antibody [1TBP18] - ChIP Grade (ab818)

This product has been referenced in:
  • Gregory BL & Cheung VG Natural variation in the histone demethylase, KDM4C, influences expression levels of specific genes including those that affect cell growth. Genome Res 24:52-63 (2014). Read more (PubMed: 24285722) »
  • Serrat N  et al. The response of secondary genes to lipopolysaccharides in macrophages depends on histone deacetylase and phosphorylation of C/EBPß. J Immunol 192:418-26 (2014). Read more (PubMed: 24307736) »

See all 79 Publications for this product

Product Wall

Application Western blot
Loading amount 60 µg
Gel Running Conditions Reduced Denaturing (10% gel)
Sample Rat Cell lysate - nuclear (airway epithelial cell)
Specification airway epithelial cell
Blocking step Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Joseph

Verified customer

Submitted Mar 07 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 15 µg
Gel Running Conditions Reduced Denaturing (5%)
Sample Mouse Cell lysate - nuclear (liver)
Specification liver
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Ms. Arlene Lim

Verified customer

Submitted Sep 04 2013

You can see this antibody used in a gel shift assay in Thompson NE et al. 2004 Protein Expression and Purification 36: 186-197. However, this publication (which must be purchased to view the entire article) refers to using a protocol for the gel shift ...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - nuclear (Hela nuclear lysate)
Loading amount 20 µg
Specification Hela nuclear lysate
Gel Running Conditions Reduced Denaturing (12)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Feb 15 2013

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We are happy to help customers find the most suitable products for their research purposes . In our review of this problem it does seem that cisplatin may affect the levels of TBP in cells while nucleolin seems...

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Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this...

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Thank you for your enquiry regarding ab818 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.
I would like to reassure you that our Abpromise applies ...

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