製品の概要

  • 製品名
    Anti-Nup153 antibody [QE5]
    Nup153 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [QE5] to Nup153
  • 特異性
    ab24700 recognises NUP153 as well as two related nuclear pore complex proteins: NUP214 and p62.
  • アプリケーション
    適用あり: Flow Cyt, WB, ICC/IF, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Human
  • 免疫原

    Full length protein: rat liver nuclear envelope proteins

  • ポジティブ・コントロール
    • In Immunocytochemistry, this antibody gave a positive signal in HepG2 cells.
  • 特記事項

    This antibody clone is manufactured by Abcam.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab24700 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
Flow Cyt Use 1-2µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB 1/1000. Detects a band of approximately 154 kDa (predicted molecular weight: 154 kDa).
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use at an assay dependent concentration. PubMed: 23423481

ターゲット情報

  • 機能
    Possible DNA-binding subunit of the nuclear pore complex (NPC). The repeat-containing domain may be involved in anchoring components of the pore complex to the pore membrane.
  • 配列類似性
    Contains 4 RanBP2-type zinc fingers.
  • ドメイン
    Contains F-X-F-G repeats.
  • 細胞内局在
    Nucleus > nuclear pore complex. Located to the terminal ring structure of the nucleoplasmic cage.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 153 kDa nucleoporin antibody
    • HNUP153 antibody
    • N153 antibody
    • NU153_HUMAN antibody
    • Nuclear pore complex protein hnup153 antibody
    • Nuclear pore complex protein Nup153 antibody
    • Nucleoporin 153kDa antibody
    • Nucleoporin Nup153 antibody
    • Nup 153 antibody
    • Nup153 antibody
    see all

画像

  • Anti-Nup153 antibody [QE5] (ab24700) at 1/1000 dilution + Human U2OS nuclear lysate at 40 µg

    Secondary
    HRP-conjugated donkey anti-mouse IgG polyclonal at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 154 kDa
    Observed band size : 160 kDa (why is the actual band size different from the predicted?)


    Exposure time : 15 minutes

    This image is courtesy of an anonymous Abreview

    See Abreview

  • Human and mouse cells treated with different concentrations of ab24700. The cells were fixed in 4% formaldehyde and permeabilized in 0.2% Triton x100 for 10 minutes at room temperature, then washed 3 times in PBS. The fixation and permeabilization was carried out in a single step. The image shows clear staining of the nuclear envalope (green). The DNA is stained with DAPI (blue).

    A: HeLa cells, ab24700 used at 4µg/ml
    B: 3T3 cells, ab24700 used at 2µg/ml
    C: 3T3 cells, ab24700 used at 4µg/ml
    D: 3T3 cells, ab24700 used at 6.6µg/ml

  • Overlay histogram showing HeLa cells stained with ab24700 (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 (ab24700, 2µg/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 IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Immunocytochemical analysis of U2OS cells, labeling Nup153 with ab24700. Cells were formaldehyde fixed, permeabilized with NP40, and blocked with 3% BSA for 1 hour at 21°C. Immunostaining with ab24700 diluted 1/100 for 12 hours at 4°C.

    See Abreview

  • ICC/IF image of ab24700 stained HepG2 cells. The cells were 4% PFA 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 (ab24700, 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.
  • Methanol fixed HeLa stained with ab24700. This antibody brilliantly highlights the nuclear membrane (green). The golgi is stained with Giantin (yellow).
  • Human and mouse cells treated with different concentrations of ab24700. The cells were fixed in 100% methanol for 20 minutes at -20°C, then washed once in PBS. The image shows clear staining of the nuclear envalope (green). The DNA is stained with DAPI (blue).

    A: HeLa cells, ab24700 used at 4µg/ml
    B: 3T3 cells, ab24700 used at 2µg/ml
    C: 3T3 cells, ab24700 used at 4µg/ml
    D: 3T3 cells, ab24700 used at 6.6µg/ml

参考文献

This product has been referenced in:
  • Mamede JI  et al. Cyclophilins and nucleoporins are required for infection mediated by capsids from circulating HIV-2 primary isolates. Sci Rep 7:45214 (2017). WB ; Human . Read more (PubMed: 28345672) »
  • Winter FR  et al. Multicolour nanoscopy of fixed and living cells with a single STED beam and hyperspectral detection. Sci Rep 7:46492 (2017). Read more (PubMed: 28417977) »

See all 28 Publications for this product

レビューと Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (U2OS)
Permeabilization
Yes - 0.5% Triton
Specification
U2OS
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formaldehyde
Username

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Verified customer

投稿 Jun 27 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (U2OS)
Permeabilization
Yes - NP40
Specification
U2OS
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

投稿 Jul 15 2015

Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing (4-12%)
Sample
Human Cell lysate - nuclear (U2OS)
Specification
U2OS
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

投稿 Apr 22 2014

Thank you for your enquiry. The antibody was raised against rat liver nuclear envelope proteins extracted in TX buffer (50 mM triethanolamine [TEA], pH 7.4, 500 mM NaCI, 0.5 % Triton X-100, 1 mM DTT, 1 mM PMSF and 1:1,000 CLAP [10 mg/ml each of chy...

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Thank you for your enquiry. I have been in touch with the source of this antibody and unfortunately I do not have a western image available at this time. I do however have soem images used to QC ths image by immunofluorescence. I have sent a follow ...

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

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