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  1. Link

    rap1-antibody-4c81-ab14404.pdf

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Epigenetics and Nuclear Signaling Chromosome Structure Telomeres
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Anti-RAP1 antibody [4c8/1] (ab14404)

  • Datasheet
  • SDS
Reviews (4)Q&A (1)References (1)

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Western blot - Anti-RAP1 antibody [4c8/1] (ab14404)
  • Immunoprecipitation - Anti-RAP1 antibody [4c8/1] (ab14404)
  • Flow Cytometry - Anti-RAP1 antibody [4c8/1] (ab14404)

Key features and details

  • Mouse monoclonal [4c8/1] to RAP1
  • Suitable for: IP, WB, Flow Cyt
  • Reacts with: Human
  • Isotype: IgG2b

You may also be interested in

Protein
Product image
Recombinant Human RAP1 protein (ab162732)
Secondary
Product image
Goat Anti-Mouse IgG H&L (HRP) (ab205719)

View more associated products

Overview

  • Product name

    Anti-RAP1 antibody [4c8/1]
    See all RAP1 primary antibodies
  • Description

    Mouse monoclonal [4c8/1] to RAP1
  • Host species

    Mouse
  • Tested applications

    Suitable for: IP, WB, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Human Rap1 protein.

  • Positive control

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

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    4c8/1
  • Myeloma

    Sp2/0-Ag14
  • Isotype

    IgG2b
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Chromosome Structure
    • Telomeres
    • Tags & Cell Markers
    • Subcellular Markers
    • Nucleus
    • Chromatin

Associated products

  • Assay kits

    • Rap1 Activation Assay Kit (ab212011)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)
  • Recombinant Protein

    • Recombinant Human RAP1 protein (ab162732)

Applications

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.

Application Abreviews Notes
IP Use a concentration of 5 µg/ml.
WB Use at an assay dependent concentration. Predicted molecular weight: 36 kDa.
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.

Target

  • Function

    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.
  • Tissue specificity

    Ubiquitous. Highly expressed.
  • Sequence similarities

    Belongs to the RAP1 family.
    Contains 1 BRCT domain.
    Contains 1 Myb-like domain.
  • Post-translational
    modifications

    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localization

    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.
  • Target information above from: UniProt accession Q9NYB0 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 54386 Human
    • Omim: 605061 Human
    • SwissProt: Q9NYB0 Human
    • Unigene: 301419 Human
    • Alternative names

      • 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

    Images

    • Western blot - Anti-RAP1 antibody [4c8/1] (ab14404)
      Western blot - Anti-RAP1 antibody [4c8/1] (ab14404)
      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
      All lanes : 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
    • Immunoprecipitation - Anti-RAP1 antibody [4c8/1] (ab14404)
      Immunoprecipitation - Anti-RAP1 antibody [4c8/1] (ab14404)
      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
    • Flow Cytometry - Anti-RAP1 antibody [4c8/1] (ab14404)
      Flow Cytometry - Anti-RAP1 antibody [4c8/1] (ab14404)
      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.

    Protocols

    • Flow cytometry protocols
    • Immunoprecipitation protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (1)

    Publishing research using ab14404? Please let us know so that we can cite the reference in this datasheet.

    ab14404 has been referenced in 1 publication.

    • Wang J  et al. Vascular Smooth Muscle Cell Senescence Promotes Atherosclerosis and Features of Plaque Vulnerability. Circulation 132:1909-19 (2015). PubMed: 26416809

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-5 of 5 Abreviews or Q&A

    Western blot abreview for Anti-RAP1 antibody [4c8/1]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 23 2016

    Western blot abreview for Anti-RAP1 antibody [4c8/1]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 23 2016

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-RAP1 antibody [4c8/1] - Telomere Marker

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted May 16 2012

    Immunocytochemistry/ Immunofluorescence abreview for Anti-RAP1 antibody [4c8/1] - Telomere Marker

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Mr. Peter Zentis

    Verified customer

    Submitted Jul 23 2010

    Question

    I am working on Drosophila Rap1. I need a good antibody to mark it's cellular localization. GTP binding domain in Drosophila and Human is very conserve. May I know which domain did you use to immunize Rap1 antibody? If it is in the conserve regions, this antibody should also work in flies theoretically.

    Read More

    Abcam community

    Verified customer

    Asked on Aug 26 2004

    Answer

    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 not, we will refund. Please also let us know if it does cross react by writing a review and in return we'll send you a USD 15.00 Amazon gift voucher.

    Read More

    Abcam Scientific Support

    Answered on Aug 27 2004

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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