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RecombinantRabMAb

Recombinant Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)

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ChIC/CUT&RUN sequencing - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
  • ChIP-sequencing - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
  • ChIP-sequencing - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
  • ChIP - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
  • Western blot - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
  • Immunoprecipitation - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
  • Flow Cytometry (Intracellular) - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
  • Immunocytochemistry/ Immunofluorescence - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
  • Immunocytochemistry/ Immunofluorescence - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
  • Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21954] to TATA binding protein TBP - ChIP Grade
  • Suitable for: ChIP-sequencing, Flow Cyt (Intra), ChIP, ChIC/CUT&RUN-seq, ICC/IF, WB, IHC-P, IP
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

APC Carrier Free HRP PE

Overview

  • Product name

    Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade
    See all TATA binding protein TBP primary antibodies

  • Description

    Rabbit monoclonal [EPR21954] to TATA binding protein TBP - ChIP Grade

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ChIP-sequencing, Flow Cyt (Intra), ChIP, ChIC/CUT&RUN-seq, ICC/IF, WB, IHC-P, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, HEK-293, HCT 116, MCF7, NIH/3T3 and C2C12 whole cell lysates; mouse and rat testis lysates. IP: HeLa whole cell lysate. IHC-P: Human testis and bladder cancer tissue; Mouse and rat testis tissues. ICC/IF: HeLa and HCT 116 cells. Flow Cyt (intra): HeLa cells. ChIP: Chromatin prepared from HeLa cells. ChIP-seq: Chromatin prepared from HeLa cells. ChIC/CUT&RUN-seq: HeLa cells.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab220788 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-sequencing
Use 4 µg for 30 µg of chromatin.
Flow Cyt (Intra)
1/500.
ChIP
Use 5 µg for 25 µg of chromatin.
ChIC/CUT&RUN-seq
Use 3µg for 105 cells.
ICC/IF
1/500.
WB
1/1000. Detects a band of approximately 45, 35 kDa (predicted molecular weight: 38 kDa).
IHC-P
1/4000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP
1/30.
Notes
ChIP-sequencing
Use 4 µg for 30 µg of chromatin.
Flow Cyt (Intra)
1/500.
ChIP
Use 5 µg for 25 µg of chromatin.
ChIC/CUT&RUN-seq
Use 3µg for 105 cells.
ICC/IF
1/500.
WB
1/1000. Detects a band of approximately 45, 35 kDa (predicted molecular weight: 38 kDa).
IHC-P
1/4000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP
1/30.

Target

  • Function

    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. 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 specificity

    Widely expressed, with levels highest in the testis and ovary.

  • Involvement in disease

    Defects 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 similarities

    Belongs to the TBP family.

  • Cellular localization

    Nucleus.
  • 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

    Images

    • ChIC/CUT&RUN sequencing - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
      ChIC/CUT&RUN sequencing - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)

      CUT&RUN was performed using the ChIC/CUT&RUN pAG-MNAse ab285373, 105 HeLa cells and 3µg of ab220788 [EPR21954]. The resulting DNA was sequenced on the NovaSeq 6000 (PE150).

      Additional screenshots of mapped reads can be downloaded here.

    • ChIP-sequencing - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
      ChIP-sequencing - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)

      Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 30 µg of chromatin and 4 µg of ab220788. ChIP DNA was sequenced on the Illumina NextSeq 500 to a depth of 30 million reads. ChIP-Seq validation performed with ChIP-Kit Transcription Factors ChIP-Seq (ab270813). 

      Additional screenshots of mapped reads can be downloaded here.

    • ChIP-sequencing - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
      ChIP-sequencing - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)

      Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 30 µg of chromatin and 4 µg of Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788). ChIP DNA was sequenced on the Illumina NextSeq 500 to a depth of 30 million reads. ChIP-Seq validation performed by Active Motif, Carlsbad, CA.

      Additional screenshots of mapped reads can be downloaded here.

    • ChIP - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
      ChIP - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)

      Chromatin was prepared from HeLa (human epithelial cell line from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol. The ChIP was performed with 25 µg of chromatin, 5 µg of ab220788 (red), and 20 µl of Protein A/G sepharose beads. 5 μg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (SYBR green approach).Primers and probes are located in the first kb of the transcribed region.

    • Western blot - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
      Western blot - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
      All lanes : Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788) at 1/1000 dilution

      Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 2 : HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate
      Lane 3 : HCT 116 (human colorectal carcinoma cell line) whole cell lysate
      Lane 4 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate
      Lane 5 : C2C12 (mouse myoblast cell line) whole cell lysate
      Lane 6 : Mouse testis lysate
      Lane 7 : Rat testis lysate
      Lane 8 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 38 kDa
      Observed band size: 35,45 kDa why is the actual band size different from the predicted?



      Exposure times : Lanes 1-7: 15 seconds, Lane 8: 3 minutes.

      Blocking and dilution buffer: 5% NFDM/TBST.

      Human TBP migrates with an approximate molecular mass of 45 kDa on SDS-PAGE (PMID:1458534; PMID: 1907890) the molecular mass observed is lower in mouse than in human samples due to a shorter sequence.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)

      Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling TATA binding protein TBP with ab220788 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining in mouse testis (PMID: 17570761; PMID: 11861477) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)

      Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling TATA binding protein TBP with ab220788 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining in rat testis (PMID: 17570761; PMID: 11861477) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)

      Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling TATA binding protein TBP with ab220788 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining in human bladder cancer (PMID: 17570761; PMID: 11861477) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)

      Immunohistochemical analysis of paraffin-embedded human testis tissue labeling TATA binding protein TBP with ab220788 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining in human testis (PMID: 17570761; PMID: 11861477) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

       

    • Immunoprecipitation - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
      Immunoprecipitation - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)

      TATA binding protein TBP was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab220788 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab220788 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

      Lane 1: HeLa whole cell lysate 10 μg (Input).
      Lane 2: ab220788 IP in HeLa whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab220788 in HeLa whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 30 seconds.

    • Flow Cytometry (Intracellular) - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
      Flow Cytometry (Intracellular) - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)

      Intracellular flow cytometric analysis of4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling TATA binding protein TBP with ab220788 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells incubated with secondary antibody only) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody. 

       

       

    • Immunocytochemistry/ Immunofluorescence - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
      Immunocytochemistry/ Immunofluorescence - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HCT 116 (human colorectal carcinoma cell line) cells labeling TATA binding protein TBP with ab220788 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in HCT 116 cells. The nuclear counter stain is DAPI (blue). 

      Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

      Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Immunocytochemistry/ Immunofluorescence - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
      Immunocytochemistry/ Immunofluorescence - Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling TATA binding protein TBP with ab220788 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in HeLa cells. The nuclear counter stain is DAPI (blue). 

      Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

      Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)
      Anti-TATA binding protein TBP antibody [EPR21954] - ChIP Grade (ab220788)

    References (1)

    Publishing research using ab220788? Please let us know so that we can cite the reference in this datasheet.

    ab220788 has been referenced in 1 publication.

    • Xue L  et al. Effect of TTLL12 on tubulin tyrosine nitration as a novel target for screening anticancer drugs in vitro. Oncol Lett 20:340 (2020). PubMed: 33123251

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