Overview

  • Product name
    Anti-Cyclin E1 antibody [HE12]
    See all Cyclin E1 primary antibodies
  • Description
    Mouse monoclonal [HE12] to Cyclin E1
  • Host species
    Mouse
  • Tested applications
    Suitable for: WBmore details
    Unsuitable for: Flow Cyt,ICC/IF,IHC-Fr or IHC-P
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human Cyclin E1.

  • Positive control
    • NIH3T3 cells
  • General notes

    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.

Properties

Applications

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.

Application Abreviews Notes
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.

  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF,IHC-Fr or IHC-P.
  • Target

    • Function
      Essential for the control of the cell cycle at the G1/S (start) transition.
    • Tissue specificity
      Highly expressed in testis and placenta. Low levels in bronchial epithelial cells.
    • Sequence similarities
      Belongs to the cyclin family. Cyclin E subfamily.
    • Post-translational
      modifications
      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.
    • Cellular localization
      Nucleus.
    • Information by UniProt
    • Database links
    • Alternative names
      • 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

    Images

    • 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.

    • 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.

    References

    This product has been referenced in:
    • He Z  et al. MiR-422a regulates cellular metabolism and malignancy by targeting pyruvate dehydrogenase kinase 2 in gastric cancer. Cell Death Dis 9:505 (2018). Read more (PubMed: 29725130) »
    • Yang S  et al. microRNA-3129 promotes cell proliferation in gastric cancer cell line SGC7901 via positive regulation of pRb. Braz J Med Biol Res 51:e6452 (2018). Read more (PubMed: 29791595) »
    See all 24 Publications for this product

    Customer reviews and Q&As

    1-10 of 19 Abreviews or Q&A

    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

    Abcam user community

    Verified customer

    Submitted Apr 22 2015

    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

    Abcam user community

    Verified customer

    Submitted Feb 25 2014

    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

    Abcam user community

    Verified customer

    Submitted Jul 30 2013

    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
    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

    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
    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

    Abcam user community

    Verified customer

    Submitted Aug 25 2011

    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

    Abcam user community

    Verified customer

    Submitted Feb 24 2011

    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

    Abcam user community

    Verified customer

    Submitted 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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