Anti-PABP 抗体 [10E10] (ab6125)

Reviews (2)Q&A (8)References (33)
Immunoprecipitation - Anti-PABP antibody [10E10] (ab6125)
  • Immunocytochemistry - Anti-PABP antibody [10E10] (ab6125)
  • Flow Cytometry - Anti-PABP antibody [10E10] (ab6125)
  • Western blot - Anti-PABP antibody [10E10] (ab6125)

Key features and details

  • Mouse monoclonal [10E10] to PABP
  • Suitable for: IP, ICC, WB, Flow Cyt
  • Reacts with: Human
  • Isotype: IgG2b

製品の概要

  • 製品名

    Anti-PABP antibody [10E10]
    PABP 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [10E10] to PABP

  • 由来種

    Mouse

  • アプリケーション

    適用あり: IP, ICC, WB, Flow Cytmore details

  • 種交差性

    交差種: Human
    非交差種: Mouse, Drosophila melanogaster

  • 免疫原

    Recombinant PABP (Human) expressed from its 1.85 kbp cDNA, NcoI to SspI.

  • 特記事項

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab6125の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
IP
Use at an assay dependent concentration.
ICC
Use at an assay dependent concentration.
WB (1)
Use at an assay dependent concentration. Predicted molecular weight: 71 kDa.
Flow Cyt
Use 1µg for 106 cells.

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

 
特記事項
IP
Use at an assay dependent concentration.
ICC
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 71 kDa.
Flow Cyt
Use 1µg for 106 cells.

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

 

ターゲット情報

  • 関連性

    The poly(A)-binding protein (PABP), which is found complexed to the 3-prime poly(A) tail of eukaryotic mRNA, is required for poly(A) shortening and translation initiation. Grange et al. (1987) isolated a melanoma cell cDNA encoding human PABP. The predicted 633-amino acid protein contains 4 repeats of an approximately 80-amino acid unit in its N-terminal half. The authors found that this repeat region is highly conserved between human and yeast PABP and is sufficient for poly(A) binding. In vitro translation of the human PABP cDNA yielded a protein with an apparent molecular mass of 73 kD by SDS-PAGE. Northern blot analysis indicated that PABP is expressed as a 2.9-kb mRNA in human melanoma cells. Gorlach et al. (1994) noted that each of the 4 repeats of PABP is a ribonucleoprotein (RNP) consensus sequence RNA-binding domain. They determined that PABP has a pI of approximately 10.3 and is a very abundant, stable protein. Immunofluorescence studies of mammalian cells indicated that PABP is located exclusively in the cytoplasm. However, using both indirect immunofluorescence and tagging of PABP1 by fusion to the green fluorescent protein (GFP), Afonina et al. (1998) demonstrated that PABP1 shuttles between the nucleus and cytoplasm. PABP1 accumulated in the nucleus when transcription was inhibited, suggesting that active transcription is required for nuclear export of PABP1.

  • 細胞内局在

    Cytoplasmic. Shuttles between the cytoplasm and the nucleus.

  • 参照データベース

    • 別名

      • PAB 1 antibody
      • PAB1 antibody
      • PABP 1 antibody
      • PABP1 antibody
      • PABPC 1 antibody
      • PABPC1 antibody
      • PABPC2 antibody
      • PABPL1 antibody
      • Poly A binding protein 1 antibody
      • Poly A binding protein cytoplasmic 1 antibody
      • poly(A) binding protein, cytoplasmic 1 antibody
      • poly(A) binding protein, cytoplasmic 2 antibody
      • Polyadenylate binding protein 1 antibody
      see all

    画像

    • Immunoprecipitation - Anti-PABP antibody [10E10] (ab6125)
      Immunoprecipitation - Anti-PABP antibody [10E10] (ab6125)
      PABP was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to PABP (ab6125) 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 ab6125.
      Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
      Band: 76kDa: PABP; 25kDa.
    • Immunocytochemistry - Anti-PABP antibody [10E10] (ab6125)
      Immunocytochemistry - Anti-PABP antibody [10E10] (ab6125)
      ICC/IF image of ab6125 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6125, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Flow Cytometry - Anti-PABP antibody [10E10] (ab6125)
      Flow Cytometry - Anti-PABP antibody [10E10] (ab6125)
      Overlay histogram showing HeLa cells stained with ab6125 (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 (ab6125, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. 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. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

    • Western blot - Anti-PABP antibody [10E10] (ab6125)
      Western blot - Anti-PABP antibody [10E10] (ab6125)

      Western Blot analysis on HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate using ab6125 at 1/100 to 1/6400 dilution. Anti-mouse IgG (whole molecule)-AP conjugate was used at the secondary at a 1/2000 dilution. Detection with BCIP/NBT substrate.
      4-12% Bis-Tris, 1X MOPS running buffer.

    参考文献 (33)

    ab6125 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab6125 は 33 報の論文で使用されています。

    • Chen D  et al. Suppressive effect of platycodin D on bladder cancer through microRNA-129-5p-mediated PABPC1/PI3K/AKT axis inactivation. Braz J Med Biol Res 54:e10222 (2021). PubMed: 33470388
    • Tauber D  et al. Modulation of RNA Condensation by the DEAD-Box Protein eIF4A. Cell 180:411-426.e16 (2020). PubMed: 31928844
    • Li S  et al. Chemosensitization of prostate cancer stem cells in mice by angiogenin and plexin-B2 inhibitors. Commun Biol 3:26 (2020). PubMed: 31942000
    • Mateu-Regué À  et al. Single mRNP Analysis Reveals that Small Cytoplasmic mRNP Granules Represent mRNA Singletons. Cell Rep 29:736-748.e4 (2019). PubMed: 31618640
    • Yang X  et al. Picornavirus 2A protease regulates stress granule formation to facilitate viral translation. PLoS Pathog 14:e1006901 (2018). PubMed: 29415027
    View all Publications for this product

    レビューと Q&A

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    1-10 of 10 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-PABP antibody [10E10]

     Below Average
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Human)
    Specification
    Human
    Fixative
    Formaldehyde
    Permeabilization
    Yes - 0.5 NP40
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C
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    Abcam user community

    Verified customer

    投稿 Jul 27 2007

    Western blot abreview for Anti-PABP antibody [10E10]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Breast epithelial cancer cell lines)
    Specification
    Breast epithelial cancer cell lines
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Read More

    Ms. Michelle Badura

    Verified customer

    投稿 Sep 02 2005

    Question

    Dear concerning,
    A user would like to use Anti-PABP antibody [10E10] (ab6125) on follicular fluids by ELISA. Could you please suggest a dilution rate for the antibody to use ELISA? Additionally, is PABP peptide (ab23710) suitable to use with PABP antibody [10E10] (ab6125) as standard?

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    Answer

    Thank you for contacting us.

    There is not a specific concentration recommended for the use of this antibody in ELISA. Optimisation is required with different dilutions in order to assess the best one for your particular conditions. We can however suggest a good starting point to begin with, for purified antibodies tested in ELISA this corresponds to 0.1ug/ml.

    Regarding the suitability of ab23710 to be used as standard with the antibody, I am afraid I cannot assure the antibody will recognize it, as its epitope has not been mapped, and we have not tested this pair in house.

    However, the peptide ab23710 has been used to raise another Anti-PABP antibody:

    https://www.abcam.com/pabp-antibody-ab21060.html

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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    Question

    What is the isotype of this antibody? It's listed as IgG2b but other sources indicate this clone is an IgG1.

    Read More
    Answer

    Thank you again for your calls and for your patience while we have looked into your enquiry. I apologize again for the delay in getting this information for you.

    My contact at the lab confirms that this antibody has an IgG2b isotype. I've looked online and there are other antibodies with the 10E10 clone number (though one is to a different protein target, Raptor). I'm not sure how to explain the discrepancy with the PABP antibody listed as an IgG1 isotype from other vendors, though most companies seem tolist this as an IgG2b clone.

    I am sorry that we don'thavemore of an explain regarding this discrepancy, but if you have any further questions or if there is anything else that we can do for you, please let me know and I'll be happy to help.

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    Question

    What is the isotype of this antibody? It's listed as IgG2a but other sources indicate this clone is an IgG1.

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    Answer

    Thank you for your calls last week and earlier this week, and I apologize for the delay getting this information for you.

    I've called over to the lab, andmy colleaguewho has more information about ab6125 has been out of the office. We should have some answers early next week.

    I apologize again that this is taking so long, and we appreciate your patience. Please let me know if you have any questions or need anything else in the meantime.

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    Question

    I have purchased the anti-PABP antibody (ab6125) and used it successfully in western blot experiments. Recently I have attempted to use it in antibody inhibition experiments with RNA and have found that the RNA is being degraded during the experiment. I was under the impression that these antibodies were highly purified and therefore should be free of ribonucleases. It would be greatly appreciated if you have knowledge about the purity of the antibodies generated (if it is lot dependent) and if abcam or any other company guarantees their antibodies to be nuclease free.

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    Answer

    Thank you for your enquiry. I have received information from our laboratory. The purification steps are not done aseptically (it's impossible) and so whilst I guess there could be some bacterially derived endonuclease, I think it unlikely as all our columns are stored in ethanol and all the tubing etc washed extensively before each run. Moreover, even if there were any endonucleases present, I would be surprised if they bound non specifically to protein G and so I wouldn't expect to see them in the end product. This is substantiated by the fact that we don't see any low (12-14Kd) MW bands in the preps when we run SDS-PAGE gels. Whilst we filter sterilize the preps prior to despatch, I guess if the antibody has subsequently been handled non-aseptically at any point, this could introduce bacteria which could produce endonuclease, however I think it unlikely. I hope this information will be useful to you. Should you require any further information, please do not hesitate to contact me.

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    Question

    Customer received 40 ul instead of 50 ul in his vial of ab6125. Lot# 135499. Order# 101326.

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    Answer

    Thank you for your phone call and I apologize for the shortage that you received. I have arranged for a free of charge replacement vial of ab6125 to be sent to your attention. It is on order# 103119 and you should receive it Tuesday. Please let me know if you have any additional questions or concerns.

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    Question

    Customer reported receiving 7 ul instead of 50 ul in her vial of ab6125. Lot# 107670. Order# 85080.

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    Answer

    Thank you for your phone call. I apologize for the shortage that you received in your vial of ab6125 and I'm sending you a replacement vial free of charge. It is on order# 86753 and you should receive this by Wednesday. Please let me know if I can be of additional assistance.

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    Question

    could you please send me the sequence of PABP that is recognized by the antibody 10E10.

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    Answer

    At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this antibody is on this datasheet. We are unable to supply the epitope sequence as this is deemed commercially sensitive information. However it may be of help to you to view the references cited at the base of the data sheet. If you need any further information please get in touch once again.

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    Question

    Customer would like to know if you have the sequence of the epitope recognized by the Mouse monoclonal [10E10] to humanPABP (ab6125).

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    Answer

    All the information that we have at this time is located on the online datasheet, and we unfortunately do not have information regarding the sequence of the epitope.

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