Recombinant
RabMAb

Recombinant Anti-Cyclin E2 antibody [E142] (ab32103)

Reviews (5)Q&A (3)References (12)

Overview

  • Product name

    Anti-Cyclin E2 antibody [E142]
    See all Cyclin E2 primary antibodies

  • Description

    Rabbit monoclonal [E142] to Cyclin E2

  • Host species

    Rabbit

  • Specificity

    ab32103 recognises Cyclin E2.

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Synthetic peptide within Human Cyclin E2 aa 350 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: O96020

  • Positive control

    • Human breast carcinoma, HeLa cells lysates, Jurkat whole cell lysate (ab7899)

  • General notes

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

Our Abpromise guarantee covers the use of ab32103 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. Predicted molecular weight: 45 kDa.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF 1/2000.

For unpurified use at 1/100 - 1/250.
Flow Cyt 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 
  • Application notes
    Is unsuitable for IP.

    • Target

    • Relevance

      The human Cyclin E2 gene encodes a 404 amino acid protein that is most closely related to Cyclin E. Cyclin E2 mRNA levels peaks at the G1 / S transition. Cyclin E2 associates with Cdk2 in a functional kinase complex that is inhibited by both p27 (Kip1) and p21 (Cip1). Cyclin E2 / Cdk2 phosphorylates histone H1 in vitro. G1 cyclin E controls the initiation of DNA synthesis by activating CDK2. Abnormally high levels of cyclin E expression have frequently been observed in human cancers. Unlike Cyclin E1, which is expressed in great majority of proliferating normal and neoplastically transformed cells, Cyclin E2 levels are low to undetectable in non transformed cells and increase significantly in neoplasm derived cells.

    • Cellular localization

      Nuclear

    • Database links

      • Alternative names

        • CCN E2 antibody
        • CCNE 2 antibody
        • CCNE2 antibody
        • CCNE2 protein antibody
        • CYC E2 antibody
        • CYCE 2 antibody
        • CYCE2 antibody
        • CyclinE2 antibody
        • G1/S specific cyclin E2 antibody
        see all

      Images

      • Western blot - Anti-Cyclin E2 antibody [E142] (ab32103)
        Western blot - Anti-Cyclin E2 antibody [E142] (ab32103)
        Anti-Cyclin E2 antibody [E142] (ab32103) at 1/500 dilution + HeLa cell lysate

        Predicted band size: 45 kDa
        Observed band size: 50 kDa
        why is the actual band size different from the predicted?

      • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E2 antibody [E142] (ab32103)
        Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E2 antibody [E142] (ab32103)

        Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling Cyclin E2 with purified ab32103 at 1/2000. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) at 1/1000 dilution was used as the secondary antibody. Nuclei counterstained with DAPI (blue).

        Secondary Only Control: PBS was used instead of the primary antibody as the negative control.

      • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E2 antibody [E142] (ab32103)
        Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E2 antibody [E142] (ab32103)
        Immunofluorescent analysis of Cyclin E2 expression in HeLa cell culture using 1/100 ab32103.
      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin E2 antibody [E142] (ab32103)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin E2 antibody [E142] (ab32103)

        Immunohistochemical analysis of Cyclin E2 expression in paraffin embedded human breast carcinoma using 1/50 ab32103.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Flow Cytometry - Anti-Cyclin E2 antibody [E142] (ab32103)
        Flow Cytometry - Anti-Cyclin E2 antibody [E142] (ab32103)
        Overlay histogram showing HeLa cells stained with ab32103 (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 (ab32103, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

      • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E2 antibody [E142] (ab32103)
        Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E2 antibody [E142] (ab32103)This image is courtesy of an abreview by Kirk Mcmanus

        ab32103 staining Cyclin E2 in the Hela cell line from Human cervix by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 22°C. Ab150081 (1/200) was used as the secondary antibody.

        See Abreview

      References

      This product has been referenced in:

      • Zhao F  et al. MicroRNA-133b suppresses bladder cancer malignancy by targeting TAGLN2-mediated cell cycle. J Cell Physiol 234:4910-4923 (2019). Read more (PubMed: 30317571) »
      • Ganguly S  et al. Pathobiology of cigarette smoke-induced invasive cancer of the renal pelvis and its prevention by vitamin C. Toxicol Rep 5:1002-1010 (2018). Read more (PubMed: 30338226) »
      See all 12 Publications for this product

      Publishing research using ab32103? Please let us know so that we can cite the reference in this datasheet.

      Customer reviews and Q&As

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      1-8 of 8 Abreviews or Q&A

      Immunocytochemistry/ Immunofluorescence abreview for Anti-Cyclin E2 antibody [E142]

       Excellent
      Abreviews
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Human Cell (HeLa)
      Permeabilization
      Yes - 0.5% Triton X100 in PBS
      Specification
      HeLa
      Fixative
      Paraformaldehyde
      Read More

      Dr. Kirk Mcmanus

      Verified customer

      Submitted Aug 25 2017

      Question

      IHC-P and chromosome spreads
      received warm and tried it, but no signal
      worked with same Ab before and it was fine
      mouse prostate tissue
      AR: Tris pH 9
      Ab: 1/100 o/n
      block: donkey serum
      2nd: OK

      Read More
      Answer

      Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.
      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 with the order number xxx. The estimated delivery date is xxx.
      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

      Western blot abreview for Anti-Cyclin E2 antibody [E142]

       Good
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Rat Tissue lysate - whole (Liver)
      Loading amount
      50 µg
      Specification
      Liver
      Gel Running Conditions
      Reduced Denaturing (12%)
      Blocking step
      Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
      Read More

      Abcam user community

      Verified customer

      Submitted Oct 28 2011

      Western blot abreview for Anti-Cyclin E2 antibody [E142]

       Good
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Mouse Cell lysate - whole cell (Hepa 1-6)
      Loading amount
      50 µg
      Specification
      Hepa 1-6
      Gel Running Conditions
      Reduced Denaturing (12%)
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
      Read More

      Abcam user community

      Verified customer

      Submitted Sep 23 2011

      Western blot abreview for Anti-Cyclin E2 antibody [E142]

       Good
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Human Cell lysate - whole cell (HepG2)
      Loading amount
      50 µg
      Specification
      HepG2
      Gel Running Conditions
      Reduced Denaturing (12)
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
      Read More

      Mrs. Sadaf Sultan

      Verified customer

      Submitted Aug 11 2011

      Western blot abreview for Anti-Cyclin E2 antibody [E142]

       Excellent
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Human Cell lysate - whole cell (HeLa cell line)
      Loading amount
      50 µg
      Specification
      HeLa cell line
      Gel Running Conditions
      Reduced Denaturing
      Blocking step
      Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
      Read More

      Dr. Shakil Ahmad

      Verified customer

      Submitted Aug 11 2011

      Question

      BATCH NUMBER 319539 ORDER NUMBER PO-00843 DESCRIPTION OF THE PROBLEM Cytoplasmic staing and heavy background SAMPLE Breast carcinoma PRIMARY ANTIBODY 1:50 for 2hours DETECTION METHOD Streptavidin HRP label POSITIVE AND NEGATIVE CONTROLS USED Pos-Human Breast carcinoma Neg- Normal rabbit IgG Fraction ANTIBODY STORAGE CONDITIONS 4 degrees FIXATION OF SAMPLE Archived formalin fixed paraffin embedded ANTIGEN RETRIEVAL Pressure cooker and pH 6.0 modified citrate PERMEABILIZATION STEP none BLOCKING CONDITIONS Avidin/Biotin blocks SECONDARY ANTIBODY Biotinylated link HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 5+ HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? No WHAT STEPS HAVE YOU ALTERED? I have taken over this project from someone else. I have no other information regarding staining parameters ADDITIONAL NOTES I am looking to steer away from the avidin/biotin system and am going to try a pH 9.0 Tris-HCL retrieval in gentle waterbath 20mins and incubate overnight at 4 degrees. There is too much cytoplasmic staining and some stromal background I must get rid of. I just wanted an idea of Abcam’s staining method for IHC so I don’t waste anymore precious tissue or reagent. And time!

      Read More
      Answer

      Thank you for your enquiry. I can confirm that this product does not require any specific protocols and the general IHC-P and antigen retrieval protocols found on our website (can also be accessed on the datasheet) will be able to help you. From the protocols you provided, I agree that you should try incubating at 4oC overnight to ensure proper and sufficient binding to the protein of interest. You can also try increasing the primary antibody dilution to 1:100 ~ 1:1000. The recommended dilution we have on the datasheet is just a guideline and optimal dilutions/concentrations should be determined by the end user. Can I also suggest running a no primary experiment? Sometimes, the problem lies with the secondary antibody and could be binding non-specifically to your sample. I hope the information above will helpful. Please do not hesitate to contact me again if you face problems when using this product.

      Read More

      Question

      I have a customer enquiring about product# ab32103, and want to know the concentration of this antibody. Could you please advise the estimated concentration for this product?

      Read More
      Answer

      Thank you for your enquiry. The approximate concentration of this antibody (ab32103) is 90ug/ml. I hope this is helpful. Please don't hesitate to contact us again if you require more information.

      Read More

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