Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR194] to Cyclin E1
- Suitable for: WB, Flow Cyt, ICC/IF
- Knockout validated
- Reacts with: Human
Product nameAnti-Cyclin E1 antibody [EPR194]
See all Cyclin E1 primary antibodies
DescriptionRabbit monoclonal [EPR194] to Cyclin E1
Tested applicationsSuitable for: WB, Flow Cyt, ICC/IFmore details
Unsuitable for: IHC-P
Species reactivityReacts with: Human
Synthetic peptide within Human Cyclin E1 (C terminal). The exact sequence is proprietary.
- HeLa whole cell lysate (ab150035), MCF7 cell lysate, JAR cell lysate, K562 cell lysate
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
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.
Storage instructionsShipped 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 bufferpH: 7.40
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.5% BSA
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab133266 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 - 1/10000. Detects a band of approximately 42-50 kDa (predicted molecular weight: 47 kDa).|
For unpurified use at 1:500.
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
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Cyclin E1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: MCF7 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab133266 observed at 47 kDa. Red - loading control, ab9484, observed at 37 kDa.
Unpurified ab133266 was shown to recognize Cyclin E1 in wild-type cells as signal was lost at the expected MW in Cyclin E1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Cyclin E1 knockout samples were subjected to SDS-PAGE. Ab133266 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 epithelial cell) cells labeling Cyclin E1 with Purified ab133266 at 1:1000 dilution (1.6μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Cyclin E1 with purified ab133266 at 1:150 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
All lanes : Anti-Cyclin E1 antibody [EPR194] (ab133266) at 1/10000 dilution (purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : JAR (Human placenta choriocarcinoma epithelial cell) whole cell lysates
Lysates/proteins at 15 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 47 kDa
Blocking and diluting buffer : 5% NFDM/TBST
All lanes : Anti-Cyclin E1 antibody [EPR194] (ab133266) at 1/1000 dilution (unpurified)
Lane 1 : HeLa cell lysate
Lane 2 : MCF7 cell lysate
Lane 3 : JAR cell lysate
Lane 4 : K562 cell lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution
Predicted band size: 47 kDa
Unpurified ab133266 staining Cyclin E1 in HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X-100 in PBS. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 22°C. ab150081, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody. Counterstained with DAPI.
ab133266 has been referenced in 3 publications.
- Zhang L & Zhang W Knockdown of NUDT21 inhibits proliferation and promotes apoptosis of human K562 leukemia cells through ERK pathway. Cancer Manag Res 10:4311-4323 (2018). PubMed: 30349365
- Yu X et al. lncRNA LINC01296 regulates the proliferation, metastasis and cell cycle of osteosarcoma through cyclin D1. Oncol Rep 40:2507-2514 (2018). PubMed: 30226542
- Lu H et al. Cell cycle-dependent phosphorylation regulates RECQL4 pathway choice and ubiquitination in DNA double-strand break repair. Nat Commun 8:2039 (2017). WB . PubMed: 29229926