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Cell Biology Cell Cycle Cyclins
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Validated using a knockout cell lineRecombinantRabMAb

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

  • Datasheet
  • SDS
Reviews (3) Submit a question References (7)

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Western blot - Anti-Cyclin E1 antibody [EPR194] (ab133266)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EPR194] (ab133266)
  • Flow Cytometry - Anti-Cyclin E1 antibody [EPR194] (ab133266)
  • Western blot - Anti-Cyclin E1 antibody [EPR194] (ab133266)
  • Western blot - Anti-Cyclin E1 antibody [EPR194] (ab133266)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EPR194] (ab133266)
  • Anti-Cyclin E1 antibody [EPR194] (ab133266)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR194] to Cyclin E1
  • Suitable for: WB, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Human

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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 & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide within Human Cyclin E1 (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

    Mouse, 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.

    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.

    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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR194
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Cyclins
    • Cell Biology
    • Cell Cycle
    • Cyclins
    • Cyclin E Family
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Cyclins
    • Cyclin E Family
    • Cancer
    • Cell cycle
    • Cyclins
    • Cyclin E family

Associated products

  • Alternative Versions

    • Anti-Cyclin E1 antibody [EPR194] - BSA and Azide free (ab208695)
    • Alexa Fluor® 647 Anti-Cyclin E1 antibody [EPR194] (ab225156)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa nuclear extract lysate (ab14655)
    • K-562 nuclear extract lysate (ab14851)
    • MCF7 nuclear extract lysate (ab14860)
  • Recombinant Protein

    • Recombinant Human Cyclin E1 protein (ab119719)

Applications

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
ICC/IF
Human
WB
Human
Application Abreviews Notes
WB (1)
1/1000 - 1/10000. Detects a band of approximately 42-50 kDa (predicted molecular weight: 47 kDa).
Flow Cyt
1/150.
ICC/IF (2)
1/1000.

For unpurified use at 1:500.

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.
    • Target information above from: UniProt accession P24864 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 898 Human
      • Omim: 123837 Human
      • SwissProt: P24864 Human
      • Unigene: 244723 Human
      • 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

      • Western blot - Anti-Cyclin E1 antibody [EPR194] (ab133266)
        Western blot - Anti-Cyclin E1 antibody [EPR194] (ab133266)

        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 - Anti-Cyclin E1 antibody [EPR194] (ab133266)
        Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EPR194] (ab133266)

        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 - Anti-Cyclin E1 antibody [EPR194] (ab133266)
        Flow Cytometry - Anti-Cyclin E1 antibody [EPR194] (ab133266)
        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).
      • Western blot - Anti-Cyclin E1 antibody [EPR194] (ab133266)
        Western blot - Anti-Cyclin E1 antibody [EPR194] (ab133266)
        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
      • Western blot - Anti-Cyclin E1 antibody [EPR194] (ab133266)
        Western blot - Anti-Cyclin E1 antibody [EPR194] (ab133266)
        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

      • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EPR194] (ab133266)
        Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EPR194] (ab133266)This image is courtesy of an Abreview submitted by Kirk McManus

        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

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

      Protocols

      • Western blot protocols
      • Immunocytochemistry & immunofluorescence protocols

      Click here to view the general protocols

      Datasheets and documents

      • SDS download

      • Datasheet download

        Download

      References (7)

      Publishing research using ab133266? Please let us know so that we can cite the reference in this datasheet.

      ab133266 has been referenced in 7 publications.

      • Wang N  et al. Yeast ß-D-glucan exerts antitumour activity in liver cancer through impairing autophagy and lysosomal function, promoting reactive oxygen species production and apoptosis. Redox Biol 32:101495 (2020). PubMed: 32171725
      • Thompson LL  et al. Reduced SKP1 Expression Induces Chromosome Instability through Aberrant Cyclin E1 Protein Turnover. Cancers (Basel) 12:N/A (2020). PubMed: 32106628
      • Velásquez ZD  et al. Besnoitia besnoiti-driven endothelial host cell cycle alteration. Parasitol Res 119:2563-2577 (2020). PubMed: 32548739
      • Moro Y  et al. In Vitro Study of the Anti-cancer Effect of Alternate-day 5-Fluorouracil in Head and Neck Cancer Cells. Anticancer Res 39:6041-6047 (2019). PubMed: 31704830
      • 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). 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). PubMed: 30226542
      • Lu H  et al. Cell cycle-dependent phosphorylation regulates RECQL4 pathway choice and ubiquitination in DNA double-strand break repair. Nat Commun 8:2039 (2017). WB . PubMed: 29229926

      Customer reviews and Q&As

      Show All Reviews Q&A
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      1-3 of 3 Abreviews

      Immunocytochemistry/ Immunofluorescence abreview for Anti-Cyclin E1 antibody [EPR194]

      Average
      Abreviews
      Abreviews
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Human Cell (DU145 Prostate cancer cell line)
      Permeabilization
      Yes - 0.1% Triton X-100
      Specification
      DU145 Prostate cancer cell line
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
      Fixative
      Paraformaldehyde
      Read More

      Dr. Dimitra Kalamida

      Verified customer

      Submitted Dec 28 2019

      Western blot abreview for Anti-Cyclin E1 antibody [EPR194]

      Excellent
      Abreviews
      Abreviews
      abreview image
      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
      Read More

      Dr. Dimitra Kalamida

      Verified customer

      Submitted Jun 21 2019

      Immunocytochemistry/ Immunofluorescence abreview for Anti-Cyclin E1 antibody [EPR194]

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

      Dr. Kirk Mcmanus

      Verified customer

      Submitted Nov 05 2014

      Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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