Anti-TATA binding protein TBP抗体-核Loading Control and ChIP Grade (ab63766)

概述

  • 产品名称
    Anti-TATA binding protein TBP抗体-核Loading Control and ChIP Grade
    参阅全部 TATA binding protein TBP 一抗
  • 描述
    兔多克隆抗体to TATA binding protein TBP -核Loading Control and ChIP Grade
  • 经测试应用
    适用于: ICC/IF, IP, ChIP, IHC-P, WBmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
    预测可用于: Chicken, Cow
  • 免疫原

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

    (Peptide available as ab64117.)

  • 阳性对照
    • WB: HeLa whole cell, HepG2 whole cell, HeLa nuclear (data not shown), HepG2 nuclear (data not shown), mouse testis tissue, NIH 3T3 whole cell and rat testis tissue lysates. IHC-P: Human breast ductal carcinoma tissue.

性能

应用

Our Abpromise guarantee covers the use of ab63766 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
ICC/IF 1/500.
IP Use at an assay dependent concentration.
ChIP Use 5 µg for 25 µg of chromatin.
IHC-P Use a concentration of 10 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 45 kDa (predicted molecular weight: 38 kDa).

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
  • 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

Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade 图像

  • Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab63766 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach for active loci and Taqman approach for inactive loci). Primers and probes are located in the first kb of the transcribed region.
  • All lanes : Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade (ab63766) at 1 µg/ml

    Lane 1 : Testis (Mouse) Tissue Lysate
    Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : Testis (Rat) Tissue Lysate
    Lane 4 : HepG2 nuclear extract lysate (ab14660)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 38 kDa
    Observed band size : 38,45 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 55 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 3 minutes
  • ab63766 staining TAT binding protein TBP in human infantile fibromatosis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% FBS/BSA for 3 hours at room temperature; antigen retrieval was by heat mediation in Tris pH9. Samples were incubated with primary antibody (1/100 in TBS + 1% BSA + 1% FBS) for 16 hours. An undiluted HRP-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview

  • TATA binding protein TBP was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Rabbit polyclonal to TATA binding protein 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, HepG2 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 ab63766.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 45kDa: TATA binding protein TBP.

  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 38 kDa
  • ab63766 (1/500) staining TATA binding protein (TBP) in HeLa cells (green). Cells were fixed with paraformaldehyde, permeabilised with 0.5% Triton X100/PBS and counterstained with DAPI (red) in order to highlight the nucleus. Please refer to abreview for further experimental details.

    See Abreview


  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 38 kDa


    Exposure time : 10 seconds
  • IHC image of TATA binding protein TBP staining in Human breast ductal carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica 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 ab63766, 10µg/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.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • All lanes : Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade (ab63766) at 1 µg/ml

    Lane 1 : HeLa Whole Cell Lysate
    Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 38 kDa
    Observed band size : 45 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 60 kDa. We are unsure as to the identity of these extra bands.

Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade (ab63766)参考文献

This product has been referenced in:
  • Tumanov S  et al. A rapid method for quantifying free and bound acetate based on alkylation and GC-MS analysis. Cancer Metab 4:17 (2016). Human . Read more (PubMed: 27594997) »
  • Sethuraman A  et al. SMARCE1 regulates metastatic potential of breast cancer cells through the HIF1A/PTK2 pathway. Breast Cancer Res 18:81 (2016). WB . Read more (PubMed: 27495308) »

See all 11 Publications for this product

Product Wall

Application
Western blot
Sample
Rat Tissue lysate - whole (liver)
Gel Running Conditions
Reduced Denaturing (8%)
Loading amount
30 µg
Specification
liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Jun 18 2015

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
Yes - tween
Specification
Brain
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 May 04 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
FBS / BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris pH 9
Sample
Human Tissue sections (Infantile fibromatosis)
Specification
Infantile fibromatosis
Permeabilization
No
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

提交于 Jul 15 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Sample
Human Cell (293T)
Specification
293T
Permeabilization
Yes - Triton X-100 1%
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

提交于 Jun 04 2014

Application
Western blot
Loading amount
5 µg
Gel Running Conditions
Reduced Denaturing (10)
Sample
Mouse Cell lysate - nuclear (mES)
Specification
mES
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 May 09 2014

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 ...

Read More

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-rea...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton X100 in PBS
Username

Dr. Kirk Mcmanus

Verified customer

提交于 Feb 24 2009

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

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