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, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Cyclin E1 aa 100-200. The exact sequence is proprietary.

  • Positive control
    • HeLa cells and 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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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 1/1000 - 1/2000. Detects a band of approximately 50 kDa (predicted molecular weight: 47 kDa).
ICC/IF 1/100 - 1/500.
Flow Cyt 1/100 - 1/1000.

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

IP 1/80.

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 [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.

  • 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.

  • Anti-Cyclin E1 antibody [EP435E] (ab33911) at 1/2000 dilution + HeLa cell lysate

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

  • 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.
  • 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.

    See 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.

    See Abreview

References

This product has been referenced in:
  • He F  et al. Effects of cullin 4B on the proliferation and invasion of human gastric cancer cells. Mol Med Rep 17:4973-4980 (2018). Read more (PubMed: 29393470) »
  • Ma Y  et al. High expression of PRPS1 induces an anti-apoptotic effect in B-ALL cell lines and predicts an adverse prognosis in Chinese children with B-ALL. Oncol Lett 15:4314-4322 (2018). Read more (PubMed: 29541198) »
See all 21 Publications for this product

Customer reviews and Q&As

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1-8 of 8 Abreviews

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.5% Triton X-100 in PBS
Specification
HeLa
Fixative
Paraformaldehyde

Dr. Kirk Mcmanus

Verified customer

Submitted Jan 16 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (ovcar4)
Gel Running Conditions
Reduced Denaturing (4-12% bis-tris)
Loading amount
10 µg
Specification
ovcar4
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Victoria Bridgeman

Verified customer

Submitted Apr 21 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: ER1 and ER2 from Leica
Sample
Human Tissue sections (ovarian cancer, TMA section)
Specification
ovarian cancer, TMA section
Permeabilization
No
Fixative
Formalin

Abcam user community

Verified customer

Submitted Mar 30 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 45 minute(s) · Concentration: 2% · Temperature: RT°C
Sample
Human Cell (HeLa cells)
Specification
HeLa cells
Permeabilization
Yes - 0.2% Triton-X100
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 15 2014

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse embryonic fibroblasts)
Loading amount
20 µg
Specification
Mouse embryonic fibroblasts
Gel Running Conditions
Reduced Denaturing (10% Tris-Gly gel)
Blocking step
LI-COR® Odyssey® Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Nov 10 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Mar 07 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
Gel Running Conditions
Reduced Denaturing (10% Tris-Gly gel)
Blocking step
LI-COR® Odyssey® Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C

Dr. Svetlana Khoronenkova

Verified customer

Submitted Aug 18 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - nuclear (HeLa)
Specification
HeLa
Gel Running Conditions
Non-reduced Denaturing (4-20% Tris-glycine)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Aug 20 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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