Recombinant Anti-Securin antibody [EPR3240] (ab79546)
![Western blot - Anti-Securin antibody [EPR3240] (ab79546)](https://www.abcam.com/ps/products/79/ab79546/Images/ab79546-365456-anti-securin-antibody-epr3240-westernblot-human.jpg "Western blot - Anti-Securin antibody [EPR3240] (ab79546)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3240] to Securin
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Securin antibody [EPR3240]
See all Securin primary antibodies -
Description
Rabbit monoclonal [EPR3240] to Securin -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human Securin aa 100-200 (N terminal). The exact sequence is proprietary.
Database link: O95997 -
Positive control
- WB: Daudi, HCT 116, HEK-293T, and HeLa cell lysates. IHC-P: Human testis and breast carcinoma tissues. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells. Jurkat 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.
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. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3240 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab79546 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) |
1/50.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
| ICC/IF | (1) |
1/50.
For unpurified use at 1/100 - 1/250. |
| WB | (2) |
1/5000 - 1/20000. Detects a band of approximately 28 kDa (predicted molecular weight: 22 kDa).
|
| IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
| Notes |
|---|
|
Flow Cyt (Intra)
1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
ICC/IF
1/50. For unpurified use at 1/100 - 1/250. |
|
WB
1/5000 - 1/20000. Detects a band of approximately 28 kDa (predicted molecular weight: 22 kDa). |
|
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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Function
Regulatory protein, which plays a central role in chromosome stability, in the p53/TP53 pathway, and DNA repair. Probably acts by blocking the action of key proteins. During the mitosis, it blocks Separase/ESPL1 function, preventing the proteolysis of the cohesin complex and the subsequent segregation of the chromosomes. At the onset of anaphase, it is ubiquitinated, conducting to its destruction and to the liberation of ESPL1. Its function is however not limited to a blocking activity, since it is required to activate ESPL1. Negatively regulates the transcriptional activity and related apoptosis activity of TP53. The negative regulation of TP53 may explain the strong transforming capability of the protein when it is overexpressed. May also play a role in DNA repair via its interaction with Ku, possibly by connecting DNA damage-response pathways with sister chromatid separation. -
Tissue specificity
Expressed at low level in most tissues, except in adult testis, where it is highly expressed. Overexpressed in many patients suffering from pituitary adenomas, primary epithelial neoplasias, and esophageal cancer. -
Sequence similarities
Belongs to the securin family. -
Developmental stage
Low level during G1 and S phases. Peaks at M phase. During anaphase, it is degraded. -
Domain
The N-terminal destruction box (D-box) acts as a recognition signal for degradation via the ubiquitin-proteasome pathway.
The TEK-boxes are required for 'Lys-11'-linked ubiquitination and facilitate the transfer of the first ubiquitin and ubiquitin chain nucleation. TEK-boxes may direct a catalytically competent orientation of the UBE2C/UBCH10-ubiquitin thiolester with the acceptor lysine residue. -
Post-translational
modificationsPhosphorylated at Ser-165 by CDK1 during mitosis.
Phosphorylated in vitro by ds-DNA kinase.
Ubiquitinated through 'Lys-11' linkage of ubiquitin moieties by the anaphase promoting complex (APC) at the onset of anaphase, conducting to its degradation. 'Lys-11'-linked ubiquitination is mediated by the E2 ligase UBE2C/UBCH10. -
Cellular localization
Cytoplasm. Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 9232 Human
- Omim: 604147 Human
- SwissProt: O95997 Human
- SwissProt: Q6IAL9 Human
- Unigene: 350966 Human
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Alternative names
- AW555095 antibody
- C87862 antibody
- Cut2 antibody
see all
Images
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Western blot - Anti-Securin antibody [EPR3240] (ab79546)All lanes : Anti-Securin antibody [EPR3240] (ab79546) at 1/1000 dilution (Purified)
Lane 1 : Daudi (Human Burkitt's lymphoma lymphoblast) whole cell lysates
Lane 2 : HCT 116 (Human colorectal carcinoma 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: 22 kDa
Observed band size: 28 kDa why is the actual band size different from the predicted? -
![Western blot - Anti-Securin antibody [EPR3240] (ab79546)](https://www.abcam.com/ps/products/79/ab79546/Images/Securin-Primary-antibodies-ab79546-1.jpg)
Western blot - Anti-Securin antibody [EPR3240] (ab79546)All lanes : Anti-Securin antibody [EPR3240] (ab79546) at 1/20000 dilution ((unpurified))
Lane 1 : Daudi (Human Burkitt's lymphoma cell line) cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 22 kDa
Observed band size: 28 kDa why is the actual band size different from the predicted?
Additional bands at: 25 kDa. We are unsure as to the identity of these extra bands. -
![Immunocytochemistry/ Immunofluorescence - Anti-Securin antibody [EPR3240] (ab79546)](https://www.abcam.com/ps/products/79/ab79546/Images/ab79546-7-anti-securin-antibody-epr3240-immunocytochemistry-hela-human.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Securin antibody [EPR3240] (ab79546)Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Securin with purified ab79546 at 1/50 dilution (4.74 µg/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). 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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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Securin antibody [EPR3240] (ab79546)](https://www.abcam.com/ps/products/79/ab79546/Images/ab79546-6-anti-securin-antibody-epr3240-immunohistochemistry-breast-carcinoma-human.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Securin antibody [EPR3240] (ab79546)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling Securin with purified ab79546 at 1/200 dilution (1.19 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody.
Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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![Flow Cytometry (Intracellular) - Anti-Securin antibody [EPR3240] (ab79546)](https://www.abcam.com/ps/products/79/ab79546/Images/ab79546-8-anti-securin-antibody-epr3240-flowcytometry-hela-human.jpg)
Flow Cytometry (Intracellular) - Anti-Securin antibody [EPR3240] (ab79546)Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Securin with purified ab79546 at 1/30 dilution (10µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilized with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr® 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).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Securin antibody [EPR3240] (ab79546)](https://www.abcam.com/ps/products/79/ab79546/Images/Securin-Primary-antibodies-ab79546-2.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Securin antibody [EPR3240] (ab79546)Unpurified ab79546 at 1/100 dilution staining Securin in paraffin-embedded human testis tissue by Immunohistochemistry.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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![Immunocytochemistry/ Immunofluorescence - Anti-Securin antibody [EPR3240] (ab79546)](https://www.abcam.com/ps/products/79/ab79546/Images/ab79546-258953-Untitled.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Securin antibody [EPR3240] (ab79546)Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling securin with unpurified ab79546 at 1/250 dilution. The cells were permeabilised with 0.1% Triton X-100. Anti-rabbit Alexa Fluor® 488 (ab150077) at 1/500 dilution was used as the secondary antibody (green). The nuclear counter stain is DAPI (blue).
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![Flow Cytometry (Intracellular) - Anti-Securin antibody [EPR3240] (ab79546)](https://www.abcam.com/ps/products/79/ab79546/Images/ab79546-258952-Untitled.jpg)
Flow Cytometry (Intracellular) - Anti-Securin antibody [EPR3240] (ab79546)Intracellular flow cytometric analysis of permeabilized Jurkat (Human T cell leukemia cell line from peripheral blood) cells using unpurified ab79546 (red) or a rabbit IgG (negative) (green).
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![Anti-Securin antibody [EPR3240] (ab79546)](https://www.abcam.com/ps/products/79/ab79546/Images/ab79546-11-benefits-of-recombinant-antibodies.png)
Anti-Securin antibody [EPR3240] (ab79546)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (25)
ab79546 has been referenced in 25 publications.
- Shindo N et al. Prolonged mitosis causes separase deregulation and chromosome nondisjunction. Cell Rep 34:108652 (2021). PubMed: 33472072
- Mecklenburg N et al. Identification of disease-relevant modulators of the SHH pathway in the developing brain. Development 148:N/A (2021). PubMed: 34463328
- Zhou C et al. The cohesin release factor Wapl interacts with Bub3 to govern SAC activity in female meiosis I. Sci Adv 6:eaax3969 (2020). PubMed: 32284991
- Konishi H et al. A tumor-specific modulation of heterogeneous ribonucleoprotein A0 promotes excessive mitosis and growth in colorectal cancer cells. Cell Death Dis 11:245 (2020). PubMed: 32303675
- Su Z et al. Analysis and Identification of Tumorigenic Targets of MicroRNA in Cancer Cells by Photoreactive Chemical Probes. Int J Mol Sci 21:N/A (2020). PubMed: 32102467