Validated using a knockout cell line
Recombinant
RabMAb

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

Overview

  • Product name

    Anti-Cyclin E1 antibody [EP435E] - BSA and Azide free
    See all Cyclin E1 primary antibodies
  • Description

    Rabbit monoclonal [EP435E] to Cyclin E1 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, Flow Cyt, ICC/IFmore 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

    Ab208696 is the carrier-free version of ab33911. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab208696 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.??

    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 ab208696 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 Use at an assay dependent concentration. Detects a band of approximately 50 kDa (predicted molecular weight: 47 kDa).
IP Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF Use at an assay dependent concentration.

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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab33911).

  • 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 data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab33911).

  • 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 data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab33911).

References

This product has been referenced in:

See all 4 Publications for this product

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