Overview

  • Product name
    Anti-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
  • Host species
    Mouse
  • Tested applications
    Suitable for: ChIP/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, Drosophila melanogaster, Silk worm
  • Immunogen

    Synthetic peptide (Human).

  • Epitope
    Within 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
  • General notes

    ab81216 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

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 concentration.
ELISA 1/1000.

We recommend Rabbit Anti-Mouse IgG H&L (Alkaline Phosphatase) (ab6729) secondary antibody.

IHC-P Use at an assay dependent concentration.
EMSA Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB 1/2000. Detects a band of approximately 38 kDa (predicted molecular weight: 38 kDa). PubMed: 23767827
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

We recommend Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879) secondary antibody

ChIP Use 5-10 µg for 25 µg of chromatin.
ICC/IF Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

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 [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
    All lanes : 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

    See Abreview

  • 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 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..

     

    See Abreview

  • 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 Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879) secondary antibody 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.

  • 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.

    See Abreview

  • 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. Rabbit Anti-Mouse IgG H&L (Alkaline Phosphatase) (ab6729) secondary antibody was used as secondary, diluted at 1/1000.

    See Abreview

  • 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



    Review by Jilin Liu submitted 19 August 2004.

References

This product has been referenced in:
  • Cai T  et al. FHL2 promotes tubular epithelial-to-mesenchymal transition through modulating ß-catenin signalling. J Cell Mol Med 22:1684-1695 (2018). WB ; Rat . Read more (PubMed: 29193729) »
  • Yin J  et al. Dkk3 dependent transcriptional regulation controls age related skeletal muscle atrophy. Nat Commun 9:1752 (2018). Read more (PubMed: 29717119) »
See all 186 Publications for this product

Customer reviews and Q&As

1-10 of 67 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Spinal cord)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer
Permeabilization
Yes - 0.2% triton x100
Specification
Spinal cord
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 27 2015

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

Joseph Cichocki

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

Ms. Arlene Lim

Verified customer

Submitted Sep 04 2013

Answer

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 assay found in Wiley SR et al. 1991 Proc Natl Acad Sci USA 89: 5814-5818 (available free of charge) with the following modifications:

A 25bp oligonucleotide containing a consensus TATA element was purchased from Promega. The reaction volumes were 20ul and contained 8ng of 5’-end-labeled 32P-labeled oligonucleotide and 10ng of human TATA Binding Protein. The reactions were incubated at 23oC for 15 minutes. For the MAb supershift assays, 40ng of MAb was added, and the reactions were allowed to incubate at 23oC for an additional 15 minutes. The reactions were subjected to electrophoresis in a nondenaturing gradient (4-15%) polyacrylamide gel on a PhastSystem for a total of 140-300Vh. The gel and buffer strips were first soaked in a solution of 1/2x TBE and 2mM MgCl2; the MgCl2 helped to stabilize the TBP/DNA complex in the absence of TFIIB.

Read More
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

Abcam user community

Verified customer

Submitted Feb 15 2013

Question
Answer

Thank you for completing our survey.

I am sorry to hear the replacement product also didn't work as expected. I would like to offer you a credit as this product is still not working as expected. Please could you confirm whether this is to your satisfaction.

I look forward to your reply.

Read More

Question
Answer

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 has caused. As requested, I have issued a free of charge replacement of ab1791 with the order number 1161045.
To check the status of the order please contact our Customer Service team and reference this number.
Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.
I wish you the best of luck with your research.

Read More

Answer

Thank you for your updates. I am very sorry to hear that even after following the suggestions, the antibody still does not perform as it is expected.
I would like to reassure you that our Abpromise applies to your complaint since you purchased this product within the guarantee period. This means that in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.
We have alternative antibodies against the same target TBP (ab51841) or against Laminin B1 (ab16048). Please could you take a look at the datasheets of these products and let me how you wish to proceed with your enquiry.
I look forward to hearing from you soon.

Read More

Answer

Thank you for contacting us.

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 affected mainly in it's distribution; however we are not specialized in this particular field and do not want to recommend a product that may be unsuitable. We encourage customers to consult the latest literature available through PubMed and other resources in order to find the most up-to-date information about their specific research interests.


I am sorry that I could not be more helpful, but I hope that the available literature in this area can provide some clarification. Please do not hesitate to contact us again with other needs or with any questions about our products.

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Question
Answer

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 has caused. As requested, I have issued a free of charge replacement with the order number 123456.
To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

1-10 of 67 Abreviews or Q&A

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

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