Anti-TATA binding protein TBP antibody [mAbcam 51841] - Nuclear Loading Control and ChIP Grade (ab51841)

Overview

  • Product name
    Anti-TATA binding protein TBP antibody [mAbcam 51841] - Nuclear Loading Control and ChIP Grade
    See all TATA binding protein TBP primary antibodies
  • Description
    Mouse monoclonal [mAbcam 51841] to TATA binding protein TBP - Nuclear Loading Control and ChIP Grade
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, ChIP, ICC/IF, Flow Cyt, ChIP/Chip, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chicken, Cow, Xenopus laevis, Chimpanzee, Zebrafish
  • Immunogen

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

    Read Abcam's proprietary immunogen policy (Peptide available as ab25711.)

  • Positive control
    • This antibody gave a positive signal in the following lysates : HeLa whole cell lysate and HeLa nuclear lysate.
  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab51841 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
WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 38 kDa (predicted molecular weight: 38 kDa).
ChIP Use 5-10 µg for 25 µg of chromatin.
ICC/IF Use a concentration of 1 µg/ml.
Flow Cyt Use 1µg for 106 cells.
ChIP/Chip Use at an assay dependent dilution.
IP Use at an assay dependent concentration.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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

  • All lanes : Anti-TATA binding protein TBP antibody [mAbcam 51841] - Nuclear Loading Control and ChIP Grade (ab51841) at 5 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Cytoplasmic Lysate at 10 µg
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 10 µg
    Lane 3 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 20 µg

    Secondary
    All lanes : Rabbit polyclonal to Mouse IgG - H&L (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size: 38 kDa
    Observed band size: 40 kDa
    why is the actual band size different from the predicted?

  • IHC image of TATA binding protein TBP staining in human breast carcinoma FFPE section, performed on a 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 ab51841, 1µg/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.
  • Chromatin was prepared from Hela 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, 5 µg of  ab51841 (blue), and 20 µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman or sybr green approach). Primers and probes are located within 1 kb of the transcription start site. 

  • Overlay histogram showing HeLa cells stained with ab51841 (red line). The cells were fixed with 80% 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 (ab51841, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • ab51841 staining TATA binding protein TBP and ab15102 staining Claudin3 in Mouse liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 30 minutes at 25°C; antigen retrieval was by heat mediation in a citrate buffer pH6. Samples were incubated with primary antibodies ab51841 and ab15102 (1/400 and 1/300 respectively in blocking buffer) for 16 hours at 4°C. A Cy3-conjugated Goat anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.

     

    See Abreview

  • ICC/IF image of ab51841 stained human HEK 293 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab51841, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa, HepG2 and MCF7 cells.

  • TBP was immunoprecipitated using 0.5mg Hela whole cell extract, 10µg of Mouse monoclonal to 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, Hela 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 ab51841.
    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.

    Band: 40kDa: TATA binding protein TBP.

References

This product has been referenced in:
  • Lakshmipathi J  et al. Identification of NFAT5 as a transcriptional regulator of the EDN1 gene in collecting duct. Am J Physiol Renal Physiol 316:F481-F487 (2019). Read more (PubMed: 30623723) »
  • Dremel SE & DeLuca NA Genome replication affects transcription factor binding mediating the cascade of herpes simplex virus transcription. Proc Natl Acad Sci U S A 116:3734-3739 (2019). Read more (PubMed: 30808759) »
See all 56 Publications for this product

Customer reviews and Q&As

1-4 of 4 Q&A

Answer



Si se pretende almacenar el anticuerpo durante largos periodos (más de una o dos semanas), lo ideal es alicuotarlo en volúmenes de trabajo, y congelarlo a -20 ºC o -80 ºC, evitando siempre ciclos de congelación / descongelación. No se recomiendan alícuotas de menos de 10ul, para evitar afectar la concentración del mismo mediante evaporación o adsorción del anticuerpo a las paredes del vial.

En cuanto al glicerol, algunos investigadores o utilizan hasta la concentración del 50% como mencionas, pero esto disminuirá el punto de congelación por debajo de -20 ºC. Salvo que lo indiquemos expresamente en la ficha técnica del producto, no consideramos necesario añadirlo al anticuerpo.

Read More

Answer

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 (˜38KDa).

For more information about loading controls you can refer to our https://www.abcam.com/index.html?pageconfig=resource&rid=11403#B5

The Abpromise® guarantee ensures that you can trust our products, and they should work in the tested species and applications stated on the datasheet, or we will offer a replacement, credit, or refund, if reported within 6 months of purchase.

I hope this helps. For more information please do not hesitate to contact us again.

Read More

Answer

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-reactivity information is stated on our datasheets, and these are updated as soon as any new information is brought to our attention.

I have checked the alignment of the immunogens with the Methanococcoides burtonii.sequence kindly provided. I am sorry this indicates only 16% alignment with all three antibodies. Therefore these antibodies are regrettablynot likely to detect the TBP from this species.

I am sorry the products are not likely to be suitable for your requirements on this occasion. However, Ihope the information is helpful. Please do not hesitate to contact me for any further advice or information.

Read More

Answer

Thank you for your enquiry.

I have confirmed the purification details of the antibodies you listed and have updated the datasheets where necessary:

ab3609 (Aurora B)
Protein G purified

ab51841 (TATA binding protein)
Protein G purified

ab37373 (goat control IgG)
Proprietary combination of ammonium sulfate precipitation, ion exchange, and/or other chromatography techniques.

ab46540 (rabbit control IgG)
Affinity purified against the antigen.

I hope this will be helpful. If you require any further information on the purification of other products, or any other questions, please do not hesitate to contact me.

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