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Anti-Cyclin E1抗体[HE12] (ab3927)

Reviews (9)Q&A (10)References (53)
Western blot - Anti-Cyclin E1 antibody [HE12] (ab3927)
  • Western blot - Anti-Cyclin E1 antibody [HE12] (ab3927)

Key features and details

  • Mouse monoclonal [HE12] to Cyclin E1
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

选择批间可重复性更高的重组抗体

Product image
Anti-Cyclin E1 antibody [EP435E] (ab33911)
  • 研究可靠 —— 各批次间结果一致且可重复
  • 长期批量供应 —— 采用重组技术，可实现快速生产
  • 首次实验即可成功 —— 经过大量验证确认了特异性
  • 符合伦理标准 —— 产品不含动物成分

概述

  • 产品名称

    Anti-Cyclin E1抗体[HE12]
    参阅全部 Cyclin E1 一抗

  • 描述

    小鼠单克隆抗体[HE12] to Cyclin E1

  • 宿主

    Mouse

  • 经测试应用

    适用于: WBmore details
    不适用于: IHC-Fr or IHC-P

  • 种属反应性

    与反应: Human

  • 免疫原

    Synthetic peptide corresponding to Human Cyclin E1.

  • 阳性对照

    • MCF7 whole cell lysate (ab3871) can be used as a positive control in WB. NIH3T3 cells

  • 常规说明

    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.

    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

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab3927于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
WB (6)
Use at an assay dependent concentration. Predicted molecular weight: 47 kDa.
说明
WB
Use at an assay dependent concentration. Predicted molecular weight: 47 kDa.
应用说明
Is unsuitable for IHC-Fr or IHC-P.

靶标

  • 功能

    Essential for the control of the cell cycle at the G1/S (start) transition.

  • 组织特异性

    Highly expressed in testis and placenta. Low levels in bronchial epithelial cells.

  • 序列相似性

    Belongs to the cyclin family. Cyclin E subfamily.

  • 翻译后修饰

    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.

  • 细胞定位

    Nucleus.
  • Information by UniProt

  • 数据库链接

    • 别名

      • 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

    图片

    • Western blot - Anti-Cyclin E1 antibody [HE12] (ab3927)
      Western blot - Anti-Cyclin E1 antibody [HE12] (ab3927)
      All lanes : Anti-Cyclin E1 antibody [HE12] (ab3927) at 1/500 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : CCNE1 (Cyclin E1) knockout HAP1 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 47 kDa



      Lanes 1 - 2: Merged signal (red and green). Green - ab3927 observed at 47 kDa. Red - loading control, ab181602, observed at 37 kDa.

      ab3927 was shown to recognize CCNE1 in wild-type HAP1 cells as signal was lost at the expected MW in CCNE1 (Cyclin E1) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and CCNE1 (Cyclin E1) knockout samples were subjected to SDS-PAGE. Ab3927 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-Cyclin E1 antibody [HE12] (ab3927)
      Western blot - Anti-Cyclin E1 antibody [HE12] (ab3927)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.

    文献 (53)

    发表研究结果有使用 ab3927？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab3927 被引用在 53 文献中.

    • Pascual J  et al. Triplet Therapy with Palbociclib, Taselisib, and Fulvestrant in PIK3CA-Mutant Breast Cancer and Doublet Palbociclib and Taselisib in Pathway-Mutant Solid Cancers. Cancer Discov 11:92-107 (2021). PubMed: 32958578
    • Kok YP  et al. Overexpression of Cyclin E1 or Cdc25A leads to replication stress, mitotic aberrancies, and increased sensitivity to replication checkpoint inhibitors. Oncogenesis 9:88 (2020). PubMed: 33028815
    • Zhu B  et al. MNX1 Promotes Malignant Progression of Cervical Cancer via Repressing the Transcription of p21cip1. Front Oncol 10:1307 (2020). PubMed: 32850410
    • Guerrero Llobet S  et al. Cyclin E expression is associated with high levels of replication stress in triple-negative breast cancer. NPJ Breast Cancer 6:40 (2020). PubMed: 32964114
    • Venugopal N  et al. The primary cilium dampens proliferative signaling and represses a G2/M transcriptional network in quiescent myoblasts. BMC Mol Cell Biol 21:25 (2020). PubMed: 32293249
    View all Publications for this product

    客户评价及客户问答

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    提交评价 提交问题

    1-10 of 19 Abreviews or Q&A

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

     Excellent
    Abreviews
    abreview image
    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

    Abcam user community

    Verified customer

    提交于 Apr 22 2015

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

     Inconclusive
    Abreviews
    abreview image
    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

    Abcam user community

    Verified customer

    提交于 Feb 25 2014

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

     Good
    Abreviews
    This product is known to not work in this application or species.
    abreview image
    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

    Abcam user community

    Verified customer

    提交于 Jul 30 2013

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

     Good
    Abreviews
    abreview image
    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

    Abcam user community

    Verified customer

    提交于 Aug 25 2011

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

     Average
    Abreviews
    abreview image
    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

    Abcam user community

    Verified customer

    提交于 Feb 24 2011

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

     Good
    Abreviews
    abreview image
    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

    Abcam user community

    Verified customer

    提交于 Feb 24 2011

    Question

    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.

    3) How long was the primary antibody incubation? overnight for 12 hours

    4) What was the blocking solution and blocking time? Odyssey Blocking Buffer for 1 hour at room temp
    .

    Read More
    Answer

    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 results.

    Read More

    Question

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

    Read More
    Answer

    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 the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

    Read More

    Question

    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.

    Read More
    Answer

    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 may not be detecting cyclin E. I would be happy to send you a replacement vial if you are interested.

    Read More

    Question

    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 repeating some of those experiments (left panel on the attached figure) and I've ordered a new vial of the same antibody (order number PR1381431). Surprisingly, I now obtain different results: a) cyclin E seems to migrate slower than in previous experiments (compare left and right panel in the attached figure). It is much closer to the 55kDa molecular marker and slower than the other cyclins. Please note the same pre-cast commercial gels are used in the 2 sets of experiments. b) while cyclin E levels increased in siEmi1 sample in the first set of experiments (right panel), in the most recent experiments there is no difference between control and siRNA sample (while the efficiency of the siRNA is comparable and the other cyclins vary in a similar manner, indicating that the experimental condition are comparable). This was observed in 3 independent experiments. Therefore, I am not sure whether the new batch of antibody is recognising cyclin E. I realise that the two batches of antibody have been ordered few years apart, but since it's a monoclonal I'd have expected them to perform in a similar manner. Do you know if anything has changed in the antibody production? If it's a problem with a specific batch, would it be possible to try an independent one of the same monoclonal?

    Read More
    Answer

    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?

    Read More

    1-10 of 19 Abreviews or Q&A

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