Customer reviews and FAQs

製品の概要

  • 製品名
    Anti-Cyclin E1 antibody [HE12]
    Cyclin E1 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [HE12] to Cyclin E1
  • アプリケーション
    適用あり: Flow Cyt, ICC/IF, IHC-P, IHC-Fr, WBmore details
  • 種交差性
    交差種: Human
  • 免疫原

    Synthetic peptide corresponding to Human Cyclin E1.

  • ポジティブ･コントロール
    • Breast carcinoma. MCF-7 cells, Hep G2, NIH3T3 cells, breast carcinoma.
  • 特記事項

    This antibody may be diluted to a titer of 1:100 - 1:200 with an enzyme-polymer detection method. The Final dilution sould be determined by the user based upon the staining conditions employed.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab3927 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 1/10 - 1/50.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF 1/50 - 1/200.
IHC-P 1/100 - 1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

This antibody may be diluted to a titer of 1:100 - 1:200 with an enzyme-polymer detection method.
IHC-Fr 1/100 - 1/200.
WB 1/500 - 1/1000. Detects a band of approximately 42 kDa (predicted molecular weight: 47 kDa).

Detects a doublet of 50 kDa and a single band of 42 kDa, see Abreviews.

ターゲット情報

  • 機能
    Essential for the control of the cell cycle at the G1/S (start) transition.
  • 組織特異性
    Highly expressed in testis and placenta. Low levels in bronchial epithelial cells.
  • 配列類似性
    Belongs to the cyclin family. Cyclin E subfamily.
  • 翻訳後修飾
    Phosphorylation of Thr-395 by GSK3 and of Ser-399 by CDK2 accelerates degradation via the ubiquitin proteasome pathway. Phosphorylated upon DNA damage, probably by ATM or ATR.
  • 細胞内局在
    Nucleus.
  • Information by UniProt
  • 参照データベース
    • 別名
      • CCNE antibody
      • Ccne1 antibody
      • CCNE1_HUMAN antibody
      • cyclin E variant ex5del antibody
      • cyclin E variant ex7del antibody
      • Cyclin E1 antibody
      • Cyclin Es antibody
      • Cyclin Et antibody
      • CyclinE antibody
      • G1/S specific cyclin E antibody
      • G1/S-specific cyclin-E1 antibody
      see all

    Anti-Cyclin E1 antibody [HE12] 画像

    • Flow Cytometry - Anti-Cyclin E1 [HE12] antibody (ab3927)
      Flow Cytometry - Anti-Cyclin E1 [HE12] antibody (ab3927)

      Overlay histogram showing MCF-7 cells stained with ab3927 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab3927, 1/10 dilution) 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 IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MCF-7 cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

    • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 [HE12] antibody (ab3927)
      Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 [HE12] antibody (ab3927)

      ICC/IF image of ab3927 stained HepG2 cells. The cells were 100% methanol fixed (5 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 (ab3927, 1/100 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) ab150113 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.

    • Western blot - Anti-Cyclin E1 [HE12] antibody (ab3927)
      Western blot - Anti-Cyclin E1 [HE12] antibody (ab3927)Image kindly provided by Tulsiram Prathapam, as part of an abreview.


      Predicted band size : 47 kDa

      Image kindly provided by Tulsiram Prathapam, as part of an abreview.

      Immunoblot analysis of Cyclin E expression in NIH/3T3 cells. Cells were transfected with the indicated plasmids and harvested 48 hr post transfection. Cyclin E expression was analysed by immunoblotting with ab3927. Immunoblot analysis of Cyclin E expression in NIH3T3 cells. Cells were transfected with the indicated plasmids and harvested 48 hr post transfection. Cyclin E expression was analysised by immunoblotting with ab3927.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin E1 [HE12] antibody (ab3927)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin E1 [HE12] antibody (ab3927)

      ab3927 staining human breast carcinoma by IHC-P.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin E1 [HE12] antibody (ab3927)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin E1 [HE12] antibody (ab3927)

      ab3927 - immunohistochemistry

      Formalin fixed paraffin embedded human breast carcinoma stained with Cyclin E, using ABC and DAB chromagen.

    Anti-Cyclin E1 antibody [HE12] (ab3927) 使用論文

    This product has been referenced in:
    • Coleman KE  et al. SENP8 limits aberrant neddylation of NEDD8 pathway components to promote cullin-RING ubiquitin ligase function. Elife 6:N/A (2017). Read more (PubMed: 28475037) »
    • Bailey ML  et al. Dependence of Human Colorectal Cells Lacking the FBW7 Tumor Suppressor on the Spindle Assembly Checkpoint. Genetics 201:885-95 (2015). WB ; Human . Read more (PubMed: 26354767) »

    See all 10 Publications for this product

    ab3927 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。

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    ab3927 の abreview を投稿する ab3927 に関する問い合わせをする

    19 Abreviews or Q&A

    10
    Votes: Highest First

    Western blot abreview for Anti-Cyclin E1 antibody [HE12]

     Excellent
    Abreviews
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HepG2 liver cells)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    50 µg
    Specification
    HepG2 liver cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More
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    Abcam user community

    Verified customer

    投稿 Apr 22 2015

    Western blot abreview for Anti-Cyclin E1 [HE12] antibody

     Inconclusive
    Abreviews
    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Reduced Denaturing (15%)
    Sample
    Mouse Cell lysate - whole cell (Mouse Pancreatic cancer)
    Specification
    Mouse Pancreatic cancer
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More
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    Abcam user community

    Verified customer

    投稿 Feb 25 2014

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Cyclin E antibody [HE12]

     Good
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
    Antigen retrieval step
    Heat mediated
    Sample
    Human Tissue sections (endometiral stromal sarcoma)
    Specification
    endometiral stromal sarcoma
    Permeabilization
    No
    Fixative
    Formaldehyde
    Read More
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    Abcam user community

    Verified customer

    投稿 Jul 30 2013

    many bands in WB
    1) What is the sample type? Metastatic Breast Cancer

    2) What lysis buffer was used? How much protein was loaded? Cell Signaling Lysis Buffer #9803S with protease and phosphatase inhibitors and with 40ug protein loaded.

    Read More

    Thank you for this additional information. One thing to try is using the 1/200 dilution for 1-2 hours room temperature, to see if you can get the target band without getting all the non-specific ones. Please let me know if this does not improve your re...

    Read More

    I'd be happy to try a replacement aliquot of the cyclin E antibody.

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab3927 with the order number 946601. To check the status of th...

    Read More

    Thanks for you reply. From previous experiments and from the observed phenotype (re-replication) cyclin E should increase in siEmi1_1. What seems strange to me is also the migration, slower than it used to be.

    Something does seem odd about the western blot data. By itself, the slightly slower migration of the cyclin E band is not that concerning. But together with the apparent insensitivity to cyclin E levels, it appears the new vial of ab3927 you received m...

    Read More

    Hi, I'm writing concerning the anti-cyclin E monoclonal antibody HE12 (cat ab3927). I have used it in the past and always obtained very good results (which also resulted in a publication, see attached figure, right panel). I am now r...

    Read More

    Thank you for your description of the issues and for including the western blot images. So you are expecting cyclin E to levels to increase due to siEmi1_1?

    Western blot abreview for Anti-Cyclin E antibody [HE12]

     Good
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLA cells)
    Loading amount
    20 µg
    Specification
    HeLA cells
    Gel Running Conditions
    Reduced Denaturing (10)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More
    abreview image
    Username

    Abcam user community

    Verified customer

    投稿 Aug 25 2011

    Western blot abreview for Anti-Cyclin E antibody [HE12]

     Average
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa cells)
    Loading amount
    20 µg
    Specification
    HeLa cells
    Treatment
    2mM double Thymidine block
    Gel Running Conditions
    Reduced Denaturing (12.5%)
    Blocking step
    Milk as blocking agent for 45 minute(s) · Concentration: 1% · Temperature: 24°C
    Read More
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    Abcam user community

    Verified customer

    投稿 Feb 24 2011

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Cyclin E antibody [HE12]

     Good
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa cells)
    Specification
    HeLa cells
    Fixative
    Methanol
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 45 minute(s) · Concentration: 1% · Temperature: 24°C
    Read More
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    Username

    Abcam user community

    Verified customer

    投稿 Feb 24 2011

    1-10 of 19 Abreviews or Q&A

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

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