Anti-RAP1抗体[4c8/1] (ab14404)

概述

  • 产品名称Anti-RAP1抗体[4c8/1]
    参阅全部 RAP1 一抗
  • 描述
    小鼠单克隆抗体[4c8/1] to RAP1
  • 经测试应用适用于: IP, WB, ELISA, Flow Cyt, IHC-P, ICC/IFmore details
  • 种属反应性
    与反应: Human
  • 免疫原

    Human Rap1 protein.

  • 阳性对照
    • In Western Blot, this antibody gave a positive signal in the following whole cell lysates: HeLa; HEK293; HepG2; A431.

性能

应用

Our Abpromise guarantee covers the use of ab14404 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
IP Use a concentration of 5 µg/ml.
WB Use at an assay dependent concentration. Predicted molecular weight: 36 kDa.
ELISA Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

ab91366 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.


ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

IHC-P 1/40.
ICC/IF Use at an assay dependent concentration.

靶标

  • 功能Acts both as a regulator of telomere function and as a transcription regulator. Involved in the regulation of telomere length and protection as a component of the shelterin complex (telosome). In contrast to other components of the shelterin complex, it is dispensible for telomere capping and does not participate in the protection of telomeres against non-homologous end-joining (NHEJ)-mediated repair. Instead, it is required to negatively regulate telomere recombination and is essential for repressing homology-directed repair (HDR), which can affect telomere length. Does not bind DNA directly: recruited to telomeric double-stranded 5'-TTAGGG-3' repeats via its interaction with TERF2. Independently of its function in telomeres, also acts as a transcription regulator: recruited to extratelomeric 5'-TTAGGG-3' sites via its association with TERF2 or other factors, and regulates gene expression. When cytoplasmic, associates with the I-kappa-B-kinase (IKK) complex and acts as a regulator of the NF-kappa-B signaling by promoting IKK-mediated phosphorylation of RELA/p65, leading to activate expression of NF-kappa-B target genes.
  • 组织特异性Ubiquitous. Highly expressed.
  • 序列相似性Belongs to the RAP1 family.
    Contains 1 BRCT domain.
    Contains 1 Myb-like domain.
  • 翻译后修饰Phosphorylated upon DNA damage, probably by ATM or ATR.
  • 细胞定位Nucleus. Cytoplasm. Chromosome. Chromosome > telomere. Associates with chromosomes, both at telomeres and in extratelomeric sites. Also exists as a cytoplasmic form, where it associates with the IKK complex.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Dopamine receptor interacting protein 5 antibody
    • Dopamine receptor-interacting protein 5 antibody
    • DRIP 5 antibody
    • DRIP5 antibody
    • hRap1 antibody
    • MGC105533 antibody
    • RAP 1 antibody
    • RAP1 homolog antibody
    • RAP1, yeast, homolog if antibody
    • RAP1, yeast, homolog of antibody
    • Repressor/activator protein 1 homolog antibody
    • TE2IP_HUMAN antibody
    • Telomeric Repeat Binding Factor 2 Interacting Protein antibody
    • Telomeric repeat-binding factor 2-interacting protein 1 antibody
    • TERF2-interacting protein antibody
    • TERF2-interacting telomeric protein 1 antibody
    • TERF2IP antibody
    • TRF2 Interacting Telomeric Protein RAP1 antibody
    • TRF2 interacting telomeric RAP1 protein antibody
    • TRF2-interacting telomeric protein 1 antibody
    • TRF2-interacting telomeric protein antibody
    see all

Anti-RAP1 antibody [4c8/1] 图像

  • ab14404 staining RAP1 in the 184A1 epithelial cell line from Human mammary glands by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with methanol/acetone (1:1), permeabilized with 1% Triton X-100 in PBS and blocked with 1% BSA for 10 minutes at 21°C. Samples were incubated with primary antibody (1/50 in PBS + 1% BSA) for 1 hour at 21°C. An undiluted Alexa Fluor®488-conjugated Donkey anti-mouse IgG polyclonal was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-RAP1 antibody [4c8/1] (ab14404) at 5 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 4 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 36 kDa
    Observed band size : 55 kDa (why is the actual band size different from the predicted?)


    Exposure time : 8 minutes
  • RAP1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to RAP1 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 ab14404.
    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
    Band: 55kDa; RAP1
  • Overlay histogram showing HeLa cells stained with ab14404 (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 (ab14404, 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 IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

Anti-RAP1 antibody [4c8/1] (ab14404)参考文献

ab14404 has not yet been referenced specifically in any publications.

Product Wall

Application Western blot
Sample African Green Monkey Cell lysate - whole cell (COS-7)
Gel Running Conditions Reduced Denaturing
Loading amount 40 µg
Specification COS-7
Blocking step BSA as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Abcam user community

Verified customer

提交于 Feb 23 2016

Application Western blot
Sample Mouse Cell lysate - whole cell (GC-1 spg)
Gel Running Conditions Reduced Denaturing (gradient 5-20%)
Loading amount 40 µg
Specification GC-1 spg
Blocking step BSA as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Feb 23 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (nerve)
Specification nerve
Fixative Formaldehyde
Antigen retrieval step Heat mediated
Permeabilization No
Blocking step antibody dilution buffer as blocking agent for 20 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 May 16 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (184A1 epithelial cell line from mamary gland)
Specification 184A1 epithelial cell line from mamary gland
Fixative Methanol and Acetone (1:1)
Permeabilization Yes - Triton X-100 1% in PBS
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
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Mr. Peter Zentis

Verified customer

提交于 Jul 23 2010

The immunogen used to generate this antibody was the Human Rap1 protein. The whole protein was used as an immunogen rather than a peptide sequence. I suggest that you purchase the antibody and determine whether it does cross react with Drosphila and if...

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