Anti-TATA binding蛋白TBP抗体[1TBP18] - ChIP Grade (ab818)

Reviews (35)Q&A (32)References (194)

概述

  • 产品名称

    Anti-TATA binding蛋白TBP抗体[1TBP18] - ChIP Grade
    参阅全部 TATA binding protein TBP 一抗

  • 描述

    小鼠单克隆抗体[1TBP18] to TATA binding蛋白TBP - ChIP Grade

  • 宿主

    Mouse

  • 经测试应用

    适用于: ChIP/Chip, ELISA, IHC-P, EMSA, IP, WB, Flow Cyt, ChIP, ICC/IF, ICC, IHC-Frmore details

  • 种属反应性

    与反应: Mouse, Rat, Human
    不与反应: Saccharomyces cerevisiae, Xenopus laevis, Drosophila melanogaster, Silk worm

  • 免疫原

    Synthetic peptide (Human).

  • 表位

    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.

  • 阳性对照

    • HeLa nuclear extract

  • 常规说明

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

    This product was changed from ascites to tissue culture supernatant on 19/12/2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions please do not hesitate to contact our scientific support team.

性能

应用

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.

应用 Ab评论 说明
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.

靶标

  • 功能

    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.

  • 组织特异性

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

  • 疾病相关

    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.

  • 序列相似性

    Belongs to the TBP family.

  • 细胞定位

    Nucleus.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • 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

图片

  • Western blot - Anti-TATA binding protein TBP antibody [1TBP18] - ChIP Grade (ab818)
    Western blot - Anti-TATA binding protein TBP antibody [1TBP18] - ChIP Grade (ab818)This image is a courtesy of Chien Hsin Lee
    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


    This image was generated using the ascites version of the product. 

    See Abreview

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

    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.

    This image was generated using the ascites version of the product. 

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TATA binding protein TBP antibody [1TBP18] - ChIP Grade (ab818)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TATA binding protein TBP antibody [1TBP18] - ChIP Grade (ab818)This image is courtesy of anonymous 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.

    This image was generated using the ascites version of the product. 

     

    See Abreview

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

    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.

    This image was generated using the ascites version of the product.

  • Immunocytochemistry/ Immunofluorescence - Anti-TATA binding protein TBP antibody [1TBP18] - ChIP Grade (ab818)
    Immunocytochemistry/ Immunofluorescence - Anti-TATA binding protein TBP antibody [1TBP18] - ChIP Grade (ab818)Image courtesy of an anonymous Abreview.

    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.

    This image was generated using the ascites version of the product. 

    See Abreview

  • ELISA - Anti-TATA binding protein TBP antibody [1TBP18] - ChIP Grade (ab818)
    ELISA - Anti-TATA binding protein TBP antibody [1TBP18] - ChIP Grade (ab818)This image is a courtesy of Anonymous 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.

    This image was generated using the ascites version of the product. 

    See Abreview

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

    This image was generated using the ascites version of the product. 

文献

This product has been referenced in:

  • Xu W  et al. Plant-derived alkaloid sinomenine potentiates glucocorticoid pharmacodynamics in mitogen-activated human peripheral blood mononuclear cells by regulating the translocation of glucocorticoid receptor. Phytother Res 33:187-196 (2019). Read more (PubMed: 30357956) »
  • Lee SH  et al. 18F-FDG positron emission tomography as a novel diagnostic tool for peripheral nerve injury. J Neurosci Methods 317:11-19 (2019). Read more (PubMed: 30684510) »
See all 194 Publications for this product

发表研究结果有使用 ab818？请让我们知道，以便我们可以引用本数据表中的参考文章。

客户评价及客户问答

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1-10 of 67 Abreviews or Q&A

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-TATA binding protein TBP antibody [1TBP18] - ChIP Grade

 Below Average
Abreviews
abreview image
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
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Abcam user community

Verified customer

提交于 Apr 27 2015

Western blot abreview for Anti-TATA binding protein TBP [1TBP18] antibody - ChIP Grade

 Excellent
Abreviews
abreview image
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
Read More

Joseph Cichocki

Verified customer

提交于 Mar 07 2014

Western blot abreview for Anti-TATA binding protein TBP antibody [1TBP18] - Nuclear Loading Control and ChIP Grade

 Excellent
Abreviews
abreview image
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
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Ms. Arlene Lim

Verified customer

提交于 Sep 04 2013

Question

I noticed for your antibody, Anti-TATA binding protein TBP antibody [1TBP18] - Nuclear Loading Control and ChIP Grade (ab818), one of the tested applications is EMSA. I can't find any protocol on your website for how to use this antibody in EMSA. Can you please give me a protocol for this? Thanks in advance for your help!

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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.

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Western blot abreview for Anti-TATA binding protein TBP antibody [1TBP18] - Nuclear Loading Control and ChIP Grade

 Excellent
Abreviews
abreview image
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
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Abcam user community

Verified customer

提交于 Feb 15 2013

Question

I received a replacement vial but it is still not wokring as expected.

Read More
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.

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Question

Yes, please!
Thank you very much!
Best

Read More
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.

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Question

Hi,

I repeated the experiment and used RIPA buffer for the nuclear fraction but still I have no signal with the anti-TBP ab818!

Do you have an other nuclear loading controle AB which I could try? Or any suggestions?

Best

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.

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Question

Hi,

I have nuclear extraction and wanted to use TBP as a loading control. However, cells were treated with cisplatin at low dose. Would it affect TBP as a loading control? If so, what do you recommend (Nucleolin would be okay?)

Thank you.

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

Customer increased work load, concentration, incubation time and developing time: no bands!

Read More
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

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