Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Cyclin E1 antibody [EPR194] (ab133266)

Overview

  • Product name

    Anti-Cyclin E1 antibody [EPR194]
    See all Cyclin E1 primary antibodies
  • Description

    Rabbit monoclonal [EPR194] to Cyclin E1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, Flow Cyt, ICC/IFmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Cyclin E1 aa 350-450 (C terminal). The exact sequence is proprietary.

  • Positive control

    • HeLa whole cell lysate (ab150035), MCF7 cell lysate, JAR cell lysate, K562 cell lysate
  • General notes

     

    A trial size is available to purchase for this antibody.

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

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab133266 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 1/1000 - 1/10000. Detects a band of approximately 42-50 kDa (predicted molecular weight: 47 kDa).
Flow Cyt 1/150.
ICC/IF 1/1000.

For unpurified use at 1:500.

  • Application notes
    Is unsuitable for 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

    • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
      Lane 2: Cyclin E1 knockout HAP1 whole cell lysate (20 µg)
      Lane 3: HeLa whole cell lysate (20 µg)
      Lane 4: MCF7 whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab133266 observed at 47 kDa. Red - loading control, ab9484, observed at 37 kDa.

      Unpurified ab133266 was shown to recognize Cyclin E1 in wild-type cells as signal was lost at the expected MW in Cyclin E1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Cyclin E1 knockout samples were subjected to SDS-PAGE. Ab133266 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Cyclin E1 with Purified ab133266 at 1:1000 dilution (1.6μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Cyclin E1 with purified ab133266 at 1:150 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
    • All lanes : Anti-Cyclin E1 antibody [EPR194] (ab133266) at 1/10000 dilution (purified)

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : JAR (Human placenta choriocarcinoma epithelial cell) whole cell lysates

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 47 kDa



      Blocking and diluting buffer : 5% NFDM/TBST
    • All lanes : Anti-Cyclin E1 antibody [EPR194] (ab133266) at 1/1000 dilution (unpurified)

      Lane 1 : HeLa cell lysate
      Lane 2 : MCF7 cell lysate
      Lane 3 : JAR cell lysate
      Lane 4 : K562 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

      Predicted band size: 47 kDa

    • Unpurified ab133266 staining Cyclin E1 in HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X-100 in PBS. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 22°C. ab150081, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody. Counterstained with DAPI.

      See Abreview

    References

    This product has been referenced in:

    • Zhang L & Zhang W Knockdown of NUDT21 inhibits proliferation and promotes apoptosis of human K562 leukemia cells through ERK pathway. Cancer Manag Res 10:4311-4323 (2018). Read more (PubMed: 30349365) »
    • Yu X  et al. lncRNA LINC01296 regulates the proliferation, metastasis and cell cycle of osteosarcoma through cyclin D1. Oncol Rep 40:2507-2514 (2018). Read more (PubMed: 30226542) »
    See all 3 Publications for this product

    Customer reviews and Q&As

    1-2 of 2 Abreviews or Q&A

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HTC116 human colorectal carcinoma)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    HTC116 human colorectal carcinoma
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

    Dr. Dimitra Kalamida

    Verified customer

    Submitted Jun 21 2019

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Permeabilization
    Yes - 0.5% Triton-X100 in PBS
    Fixative
    Paraformaldehyde

    Dr. Kirk Mcmanus

    Verified customer

    Submitted Nov 05 2014

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