Recombinant Anti-Cyclin E1 antibody [EPR194] (ab133266)
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: Flow Cyt (Intra), WB, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Cyclin E1 antibody [EPR194]
See all Cyclin E1 primary antibodies -
Description
Rabbit monoclonal [EPR194] to Cyclin E1 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, ICC/IFmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa whole cell lysate (ab150035), MCF7 cell lysate, JAR cell lysate, K562 cell lysate and Wild-type HAP1 whole cell lysate. ICC/IF: HeLa cells Flow Cyt (Intra): MCF7 cells
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General notes
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped 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 buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR194 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
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.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/150.
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WB | (1) |
1/1000 - 1/10000. Detects a band of approximately 42-50 kDa (predicted molecular weight: 47 kDa).
|
ICC/IF | (2) |
1/1000.
For unpurified use at 1:500. |
Notes |
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Flow Cyt (Intra)
1/150. |
WB
1/1000 - 1/10000. Detects a band of approximately 42-50 kDa (predicted molecular weight: 47 kDa). |
ICC/IF
1/1000. For unpurified use at 1:500. |
Target
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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
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. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 898 Human
- Omim: 123837 Human
- SwissProt: P24864 Human
- Unigene: 244723 Human
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Alternative names
- CCNE antibody
- Ccne1 antibody
- CCNE1_HUMAN antibody
see all
Images
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All lanes : Anti-Cyclin E1 antibody [EPR194] (ab133266) at 1/1000 dilution
Lane 1 : Wild-type MCF7 cell lysate
Lane 2 : CCNE1 knockout MCF7 cell lysate
Lane 3 : HT-29 cell lysate
Lane 4 : PC-3 cell lysate
Lysates/proteins at 20 µg per lane.
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 47 kDa
Observed band size: 47 kDaWestern blot: Anti-CCNE1 antibody [EPR194] (ab133266) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab133266 was shown to bind specifically to CCNE1. A band was observed at 47 kDa in wild-type MCF7 cell lysates with no signal observed at this size in CCNE1 knockout cell line ab286303 (knockout cell lysate AB300211). To generate this image, wild-type and CCNE1 knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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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.
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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.
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Intracellular 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).
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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.
Secondary
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.
Secondary
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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (17)
ab133266 has been referenced in 17 publications.
- Wu Q et al. Lnc-hipk1 inhibits mouse adipocyte apoptosis as a sponge of miR-497. Biofactors 48:135-147 (2022). PubMed: 34856026
- Auwercx J et al. TRPM7 Modulates Human Pancreatic Stellate Cell Activation. Cells 11:N/A (2022). PubMed: 35883700
- Zhang HR et al. LncRNA-cCSC1 promotes cell proliferation of colorectal cancer through sponging miR-124-3p and upregulating CD44. Biochem Biophys Res Commun 557:228-235 (2021). PubMed: 33887588
- Qin Y et al. Overexpressed lncRNA AC068039.4 Contributes to Proliferation and Cell Cycle Progression of Pulmonary Artery Smooth Muscle Cells Via Sponging miR-26a-5p/TRPC6 in Hypoxic Pulmonary Arterial Hypertension. Shock 55:244-255 (2021). PubMed: 33026218
- Lee JH et al. CDK2 phosphorylation of Werner protein (WRN) contributes to WRN's DNA double-strand break repair pathway choice. Aging Cell 20:e13484 (2021). PubMed: 34612580