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    products/primary-antibodies/cyclin-e1-antibody-ep435e-ab33911.pdf

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Cell Biology Cell Cycle Cyclins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Cyclin E1 antibody [EP435E] (ab33911)

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  • SDS
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Western blot - Anti-Cyclin E1 antibody [EP435E] (ab33911)
  • Western blot - Anti-Cyclin E1 antibody [EP435E] (ab33911)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EP435E] (ab33911)
  • Immunoprecipitation - Anti-Cyclin E1 antibody [EP435E] (ab33911)
  • Western blot - Anti-Cyclin E1 antibody [EP435E] (ab33911)
  • Flow Cytometry (Intracellular) - Anti-Cyclin E1 antibody [EP435E] (ab33911)
  • Flow Cytometry (Intracellular) - Anti-Cyclin E1 antibody [EP435E] (ab33911)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EP435E] (ab33911)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EP435E] (ab33911)
  • Anti-Cyclin E1 antibody [EP435E] (ab33911)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP435E] to Cyclin E1
  • Suitable for: WB, ICC/IF, IP, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 Carrier Free

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Recombinant Human Cyclin E1 protein (ab119719)
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Anti-Cyclin E1 antibody [EP435E] - BSA and Azide free (ab208696)
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Overview

  • Product name

    Anti-Cyclin E1 antibody [EP435E]
    See all Cyclin E1 primary antibodies
  • Description

    Rabbit monoclonal [EP435E] to Cyclin E1
  • Host species

    Rabbit
  • Specificity

    This antibody recognises Cyclin E1. It is predicted to detect the splice isoform 2 based on sequence analysis.
  • Tested applications

    Suitable for: WB, ICC/IF, IP, Flow Cyt (Intra)more details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HAP1 and HeLa cell lysates, Human testis and placenta tissue lysates IP: HeLa cell lysate Flow Cyt (intra): HeLa and MCF7 Cells ICC/IF: HeLa cells
  • General notes

    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.

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

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: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP435E
  • 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

    • Alexa Fluor® 647 Anti-Cyclin E1 antibody [EP435E] (ab194069)
    • Anti-Cyclin E1 antibody [EP435E] - BSA and Azide free (ab208696)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

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

    • HeLa whole cell lysate (ab29545)
  • Recombinant Protein

    • Recombinant Human Cyclin E1 protein (ab119719)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab33911 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 (4)
1/1000. Detects a band of approximately 50 kDa (predicted molecular weight: 47 kDa).
ICC/IF (2)
1/100 - 1/500.
IP (1)
1/30.
Flow Cyt (Intra)
1/30.

For unpurified use 1/100 - 1/1000.  ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Notes
WB
1/1000. Detects a band of approximately 50 kDa (predicted molecular weight: 47 kDa).
ICC/IF
1/100 - 1/500.
IP
1/30.
Flow Cyt (Intra)
1/30.

For unpurified use 1/100 - 1/1000.  ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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 [EP435E] (ab33911)
      Western blot - Anti-Cyclin E1 antibody [EP435E] (ab33911)
      All lanes : Anti-Cyclin E1 antibody [EP435E] (ab33911) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 cell lysate
      Lane 2 : CCNE1 knockout HAP1 cell lysate
      Lane 3 : Jurkat cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 47 kDa
      Observed band size: 47 kDa



      False colour image of Western blot: Anti-Cyclin E1 antibody [EP435E] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab33911 was shown to bind specifically to Cyclin E1. A band was observed at 47 kDa in wild-type HAP1 cell lysates with no signal observed at this size in CCNE1 knockout cell line. To generate this image, wild-type and CCNE1 knockout HAP1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

    • Western blot - Anti-Cyclin E1 antibody [EP435E] (ab33911)
      Western blot - Anti-Cyclin E1 antibody [EP435E] (ab33911)
      All lanes : Anti-Cyclin E1 antibody [EP435E] (ab33911) at 1/1000 dilution

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

      Lysates/proteins at 40 µg per lane.

      Predicted band size: 47 kDa



      Lanes 1 - 2: Merged signal (red and green). Green - ab33911 observed at 47 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab33911 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. Ab33911 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.

      This image was generated using the unpurified format of the antibody.

    • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EP435E] (ab33911)
      Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EP435E] (ab33911)

      Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labeling Cyclin E1 (green) with purified ab33911 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Nuclei were counterstained with DAPI (blue).

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

    • Immunoprecipitation - Anti-Cyclin E1 antibody [EP435E] (ab33911)
      Immunoprecipitation - Anti-Cyclin E1 antibody [EP435E] (ab33911)

      Purified ab33911 at 1/30 dilution (2ug) immunoprecipitating Cyclin E1 in HeLa whole cell lysate.
      Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate (10µg)
      Lane 2 (+): ab33911 + HeLa whole cell lysate.
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab33911 in HeLa whole cell lysate.
      VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000) was used for Western blotting.
      Blocking Buffer and concentration: 5% NFDM/TBST.
      Diluting buffer and concentration: 5% NFDM/TBST.
      Observed band size: 50 kDa

    • Western blot - Anti-Cyclin E1 antibody [EP435E] (ab33911)
      Western blot - Anti-Cyclin E1 antibody [EP435E] (ab33911)
      Lane 1 : Anti-Cyclin E1 antibody [EP435E] (ab33911) at 1/1000 dilution (Purified)
      Lanes 2-3 : Anti-Cyclin E1 antibody [EP435E] (ab33911) at 1/1000 dilution

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : Human testis lysates
      Lane 3 : Human placenta lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

      Predicted band size: 47 kDa
      Observed band size: 50 kDa why is the actual band size different from the predicted?



      Cyclin E1 is highly expressed in testis and placenta which is described in PMID: 9840943.

      Blocking/Diluting buffer: 5% NFDM/TBST.

    • Flow Cytometry (Intracellular) - Anti-Cyclin E1 antibody [EP435E] (ab33911)
      Flow Cytometry (Intracellular) - Anti-Cyclin E1 antibody [EP435E] (ab33911)

      Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Cyclin E1 with Purified ab33911 at 1/30 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

       

    • Flow Cytometry (Intracellular) - Anti-Cyclin E1 antibody [EP435E] (ab33911)
      Flow Cytometry (Intracellular) - Anti-Cyclin E1 antibody [EP435E] (ab33911)

      Overlay histogram showing MCF7 cells stained with ab33911 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab33911, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in MCF7 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions. 

      This image was generated using the unpurified format of the antibody. 

       

       

    • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EP435E] (ab33911)
      Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EP435E] (ab33911)This image is courtesy of an Abreview submitted by Kirk McManus, Univ. of Manitoba/Cancer Care MICB.

      Immunocytochemistry/Immunofluorescence analysis of HeLa cells labeling Cyclin E1 with ab33911 at 1/500 dilution. Cells were fixed in paraformaldehyde and permeabilized with 0.5% Triton X-100 in PBS. Staining with ab33911 at 1/500 was carried out for 1 hour at 22°C in PBS buffer. ab150081, a Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody, was used at 1/200 dilution. DAPI was used to counterstain.

      This image was generated using the unpurified format of the antibody.

      See Abreview

    • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EP435E] (ab33911)
      Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EP435E] (ab33911)This image is courtesy of an anonymous Abreview

      ab33911 staining Cyclin E1 in HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.2% Triton X-100 and blocked with 2% BSA for 45 minutes at room temperature. Samples were incubated with primary antibody (1/300 in PBS + 2% BSA) for 14 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

      This image was generated using the unpurified format of the antibody.

      See Abreview

    • Anti-Cyclin E1 antibody [EP435E] (ab33911)
      Anti-Cyclin E1 antibody [EP435E] (ab33911)

    Protocols

    • Flow cytometry protocols
    • Immunoprecipitation protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (91)

    Publishing research using ab33911? Please let us know so that we can cite the reference in this datasheet.

    ab33911 has been referenced in 91 publications.

    • Wang QZ  et al. Exosome-derived miR-196b-5p facilitates intercellular interaction in infantile hemangioma via down-regulating CDKN1B. Ann Transl Med 9:394 (2021). PubMed: 33842615
    • Hao L  et al. ALKBH5-mediated m6A demethylation of FOXM1 mRNA promotes progression of uveal melanoma. Aging (Albany NY) 13:4045-4062 (2021). PubMed: 33428593
    • Shen H  et al. Curcumin analog B14 has high bioavailability and enhances the effect of anti-breast cancer cells in vitro and in vivo. Cancer Sci 112:815-827 (2021). PubMed: 33316116
    • Xu X  et al. SPTBN1 Prevents Primary Osteoporosis by Modulating Osteoblasts Proliferation and Differentiation and Blood Vessels Formation in Bone. Front Cell Dev Biol 9:653724 (2021). PubMed: 33816505
    • Chen L  et al. Long Non-coding RNA SENP3-EIF4A1 Functions as a Sponge of miR-195-5p to Drive Triple-Negative Breast Cancer Progress by Overexpressing CCNE1. Front Cell Dev Biol 9:647527 (2021). PubMed: 33791304
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Immunoprecipitation abreview for Anti-Cyclin E antibody [EP435E]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Sample
    Human Cell lysate - whole cell (HeLa cells)
    Total protein in input
    200 µg
    Specification
    HeLa cells
    Immuno-precipitation step
    Protein A/G
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Mar 07 2011

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