Product nameAnti-Cyclin E1 antibody [EP435E]
See all Cyclin E1 primary antibodies
DescriptionRabbit monoclonal [EP435E] to Cyclin E1
SpecificityThis antibody recognises Cyclin E1. It is predicted to detect the splice isoform 2 based on sequence analysis.
Tested applicationsSuitable for: WB, ICC/IF, Flow Cyt, IPmore details
Species reactivityReacts with: Human
Synthetic peptide within Human Cyclin E1 aa 100-200. The exact sequence is proprietary.
- HeLa cells and cell lysate.
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.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol, 0.05% BSA
Concentration information loading...
- Anti-Cyclin E1 antibody [EP435E] (Alexa Fluor® 488) (ab194068)
- Anti-Cyclin E1 antibody [EP435E] (Alexa Fluor® 647) (ab194069)
- Anti-Cyclin E1 antibody [EP435E] (HRP) (ab194070)
- Anti-Cyclin E1 antibody [EP435E] - BSA and Azide free (ab208696)
- Anti-Cyclin E1 antibody [EP435E] (Alexa Fluor® 555) (ab215215)
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.
|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.
FunctionEssential for the control of the cell cycle at the G1/S (start) transition.
Tissue specificityHighly expressed in testis and placenta. Low levels in bronchial epithelial cells.
Sequence similaritiesBelongs to the cyclin family. Cyclin E subfamily.
modificationsPhosphorylation 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.
- Information by UniProt
- CCNE antibody
- Ccne1 antibody
- CCNE1_HUMAN antibody
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.
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 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) »