Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

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 - BSA and Azide free
  • Suitable for: WB, IP, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Human

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. 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: HeLa and MCF7 Cells ICC/IF: HeLa cells

  • 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

    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.

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-Cyclin E1 antibody [EP435E] - BSA and Azide free (ab208696)
    Western blot - Anti-Cyclin E1 antibody [EP435E] - BSA and Azide free (ab208696)
    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) (unpurified).

  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EP435E] - BSA and Azide free (ab208696)
    Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EP435E] - BSA and Azide free (ab208696)

    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).

  • Western blot - Anti-Cyclin E1 antibody [EP435E] - BSA and Azide free (ab208696)
    Western blot - Anti-Cyclin E1 antibody [EP435E] - BSA and Azide free (ab208696)
    All lanes : Anti-Cyclin E1 antibody [EP435E] (ab33911) at 1/1000 dilution (Purified)

    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.

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

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

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

  • Flow Cytometry - Anti-Cyclin E1 antibody [EP435E] - BSA and Azide free (ab208696)
    Flow Cytometry - Anti-Cyclin E1 antibody [EP435E] - BSA and Azide free (ab208696)
    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).
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab33911).
  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EP435E] - BSA and Azide free (ab208696)
    Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EP435E] - BSA and Azide free (ab208696) This image is courtesy of an Abreview submitted by Kirk McManus.

    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) (unpurified).

  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EP435E] - BSA and Azide free (ab208696)
    Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EP435E] - BSA and Azide free (ab208696) 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 data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab33911) (unpurified).

  • Flow Cytometry - Anti-Cyclin E1 antibody [EP435E] - BSA and Azide free (ab208696)
    Flow Cytometry - Anti-Cyclin E1 antibody [EP435E] - BSA and Azide free (ab208696)

    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) (unpurified).

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

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