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
  • アプリケーション適用あり: ELISA, WB, IP, ICC/IF, Flow Cytmore details
  • 種交差性
    交差種: Rabbit, Chicken, Human, Xenopus laevis
    非交差種: Mouse, Drosophila melanogaster
  • 免疫原

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

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab6125 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ELISA Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 70 kDa (predicted molecular weight: 71 kDa).
IP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration. PubMed: 17977970
Flow Cyt Use 1µg for 106 cells.

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

Anti-PABP antibody [10E10] 画像

  • All lanes : Anti-PABP antibody [10E10] (ab6125) at 1/2000 dilution

    Lane 1 : 10ug of protein from MCF10A cells transfected with negative control siRNA.
    Lane 2 : 10ug of protein from MCF10A cells transfected with PABP specific siRNA.
    Lane 3 : 10 ug of protein from MCF10A cells transfected with PABP specific siRNA.

    Secondary
    Goat anti-mouse 1/10000

    Predicted band size : 71 kDa
    Observed band size : 70 kDa (why is the actual band size different from the predicted?)

    This image is courtesy of an abreview submitted by Francisco Ramirez-Valle

    The cells were lysed with NP40 buffer with protease inhibitor cocktail, 10 micrograms of protein were separated by SDS-PAGE and transferred to PVDF membrane, blocked and blotted for two hours with PABP antibody in TBST, washed three times, secondary antibody for 1 hr (goat anti-mouse, Amersham, 1:10000).
  • 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.
  • 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.

  • 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.

Anti-PABP antibody [10E10] (ab6125) 使用論文

This product has been referenced in:
  • Backlund M  et al. Endoplasmic reticulum stress increases AT1R mRNA expression via TIA-1-dependent mechanism. Nucleic Acids Res 44:3095-104 (2016). ICC/IF . Read more (PubMed: 26681690) »
  • Tahiri-Alaoui A  et al. Poly(A) binding protein 1 enhances cap-independent translation initiation of neurovirulence factor from avian herpesvirus. PLoS One 9:e114466 (2014). Read more (PubMed: 25503397) »

See all 17 Publications for this product

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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 how...

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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
Username

Abcam user community

Verified customer

投稿 Jul 27 2007

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 k...

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Abcam guarantees this product to work in the species/application used in this Abreview.
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%
Username

Ms. Michelle Badura

Verified customer

投稿 Sep 02 2005

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 ...

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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. H...

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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.

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