Recombinant Anti-Cyclin E2 antibody [EP454Y] (ab40890)
![Western blot - Anti-Cyclin E2 antibody [EP454Y] (ab40890)](https://www.abcam.com/ps/products/40/ab40890/Images/ab40890-369829-anti-cyclin-e2-antibody-ep454y-western-blot-hap1-wt-hap1-mapk6-knockout-hap1-ccne2-knockout-mcf7.jpg "Western blot - Anti-Cyclin E2 antibody [EP454Y] (ab40890)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP454Y] to Cyclin E2
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Cyclin E2 antibody [EP454Y]
See all Cyclin E2 primary antibodies -
Description
Rabbit monoclonal [EP454Y] to Cyclin E2 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human Cyclin E2 aa 100-200. The exact sequence is proprietary.
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Positive control
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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.
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.
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: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP454Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab40890 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 |
|---|---|---|
| Flow Cyt (Intra) |
1/20.
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| WB |
1/1000 - 1/10000. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).
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| IHC-P |
1/100 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| ICC/IF | (1) |
1/50 - 1/250.
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| IP |
1/20 - 1/50.
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| Notes |
|---|
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Flow Cyt (Intra)
1/20. |
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WB
1/1000 - 1/10000. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa). |
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IHC-P
1/100 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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ICC/IF
1/50 - 1/250. |
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IP
1/20 - 1/50. |
Target
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Relevance
The human Cyclin E2 gene encodes a 404 amino acid protein that is most closely related to Cyclin E. Cyclin E2 mRNA levels peaks at the G1 / S transition. Cyclin E2 associates with Cdk2 in a functional kinase complex that is inhibited by both p27 (Kip1) and p21 (Cip1). Cyclin E2 / Cdk2 phosphorylates histone H1 in vitro. G1 cyclin E controls the initiation of DNA synthesis by activating CDK2. Abnormally high levels of cyclin E expression have frequently been observed in human cancers. Unlike Cyclin E1, which is expressed in great majority of proliferating normal and neoplastically transformed cells, Cyclin E2 levels are low to undetectable in non transformed cells and increase significantly in neoplasm derived cells. -
Cellular localization
Nuclear -
Database links
- Entrez Gene: 9134 Human
- Omim: 603775 Human
- SwissProt: O96020 Human
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Alternative names
- CCN E2 antibody
- CCNE 2 antibody
- CCNE2 antibody
see all
Images
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Western blot - Anti-Cyclin E2 antibody [EP454Y] (ab40890)All lanes : Anti-Cyclin E2 antibody [EP454Y] (ab40890) at 1/10000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : CCNE2 knockout HAP1 whole cell lysate
Lane 3 : MCF7 whole cell lysate
Lane 4 : HeLa whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab40890 observed at 45 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab40890 was shown to react with CCNE2 in HAP1 wild-type cells in Western blot. Loss of signal was observed when CCNE2 knockout sample was used. HAP1 wild-type and CCNE2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with ab40890 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 10000 dilution and a 1 in 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® 800CW) preabsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin E2 antibody [EP454Y] (ab40890)](https://www.abcam.com/ps/products/40/ab40890/Images/ab40890-5-anti-cyclin-e2-antibody-ep454y-immunohistochemistry-breast-carcinoma-human.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin E2 antibody [EP454Y] (ab40890)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling Cyclin E2 with purified ab40890 at 1/100 dilution (1.41 µg/ml). Heat mediated antigen retrieval was performed Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. -
![Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E2 antibody [EP454Y] (ab40890)](https://www.abcam.com/ps/products/40/ab40890/Images/ab40890-6-anti-cyclin-e2-antibody-ep454y-immunocytochemistry-hela-human.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E2 antibody [EP454Y] (ab40890)Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Cyclin E2 with purified ab40890 at 1:50 dilution (2.8 µg/ml). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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![Western blot - Anti-Cyclin E2 antibody [EP454Y] (ab40890)](https://www.abcam.com/ps/products/40/ab40890/Images/ab40890-351347-anti-cyclin-e2-antibody-ep454y-westernblot-jurkat-human.jpg)
Western blot - Anti-Cyclin E2 antibody [EP454Y] (ab40890)Anti-Cyclin E2 antibody [EP454Y] (ab40890) at 1/1000 dilution (Purified) + Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 50 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted? -
![Immunoprecipitation - Anti-Cyclin E2 antibody [EP454Y] (ab40890)](https://www.abcam.com/ps/products/40/ab40890/Images/ab40890-7-anti-cyclin-e2-antibody-ep454y-immunoprecipitation-jurkat-human.jpg)
Immunoprecipitation - Anti-Cyclin E2 antibody [EP454Y] (ab40890)ab40890 (purified) at 1/20 dilution (1 µg) immunoprecipitating Cyclin E2 in Jurkat whole cell lysate.
Lane 1 (input): Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10 µg
Lane 2 (+): ab40890 & Jurkat whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab40890 in Jurkat whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin E2 antibody [EP454Y] (ab40890)](https://www.abcam.com/ps/products/40/ab40890/Images/ab40890-54209-anti-cyclin-e2-antibody-ep454y-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin E2 antibody [EP454Y] (ab40890)ab40890 (unpurified) at 1/250 staining human breast carcinoma by immunohistochemistry, paraffin-embedded tissue.
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![Flow Cytometry (Intracellular) - Anti-Cyclin E2 antibody [EP454Y] (ab40890)](https://www.abcam.com/ps/products/40/ab40890/Images/ab40890-280374-anti-cyclin-e2-antibody-ep454y-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-Cyclin E2 antibody [EP454Y] (ab40890)Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Cyclin E2 (right) with purified ab40890 at a 1/20 dilution. Cells were fixed with 90% ethanol and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluorr® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution.
Left panel - Rabbit monoclonal IgG (ab172730).
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![Anti-Cyclin E2 antibody [EP454Y] (ab40890)](https://www.abcam.com/ps/products/40/ab40890/Images/ab40890-10-benefits-of-recombinant-antibodies.png)
Anti-Cyclin E2 antibody [EP454Y] (ab40890)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (22)
ab40890 has been referenced in 22 publications.
- Kaminska K et al. Distinct mechanisms of resistance to fulvestrant treatment dictate level of ER independence and selective response to CDK inhibitors in metastatic breast cancer. Breast Cancer Res 23:26 (2021). PubMed: 33602273
- Huang S et al. Obg-like ATPase 1 (OLA1) overexpression predicts poor prognosis and promotes tumor progression by regulating P21/CDK2 in hepatocellular carcinoma. Aging (Albany NY) 12:3025-3041 (2020). PubMed: 32045367
- Lee C et al. Cyclin E2 Promotes Whole Genome Doubling in Breast Cancer. Cancers (Basel) 12:N/A (2020). PubMed: 32823571
- Chen R et al. Cx43 and AKAP95 regulate G1/S conversion by competitively binding to cyclin E1/E2 in lung cancer cells. Thorac Cancer 11:1594-1602 (2020). PubMed: 32338437
- Lorenzon I et al. Identification and Characterization of a New Platinum-Induced TP53 Mutation in MDAH Ovarian Cancer Cells. Cells 9:N/A (2019). PubMed: 31877751