Recombinant Anti-PTEN (phospho T366) antibody [EP229] (ab109454)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP229] to PTEN (phospho T366)
- Suitable for: WB, IHC-P, Dot blot
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-PTEN (phospho T366) antibody [EP229]
See all PTEN primary antibodies -
Description
Rabbit monoclonal [EP229] to PTEN (phospho T366) -
Host species
Rabbit -
Specificity
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
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Tested applications
Suitable for: WB, IHC-P, Dot blotmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa (Human cervix adenocarcinoma epithelial cell), NIH/3T3 (Mouse embryonic fibroblast), and C6 (Rat glial tumor glial cell) whole cell lysate untreated, treated with 100ng/ml Calyculin A for 30 minutes, and ted with 100ng/ml Calyculin A for 30 minutes whole cell lysate then the membrane was incubated with Alkaline phosphatase. IHC-P: Human breast ductal carcinoma tissue and prostate cancer sections.
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General notes
This product has switched from a hybridoma to recombinant production method on 9th June 2023.
PTEN is a protein implicated in several disease, including certain cancers and neurological diseases. PTEN is expressed ubiquitously throughout the body and acts as a phosphatase to dephosphorylate phosphatidylinositol (3,4,5)-trisphosphate. This is important in the inhibition of the Akt signalling pathway, which plays an important role in regulating cellular behaviours such as cell growth, survival, and migration.
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.
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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP229 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab109454 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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WB |
1/10000. Detects a band of approximately 54 kDa (predicted molecular weight: 47 kDa).
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IHC-P |
1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. |
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Dot blot |
1/10000.
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Notes |
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WB
1/10000. Detects a band of approximately 54 kDa (predicted molecular weight: 47 kDa). |
IHC-P
1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. |
Dot blot
1/10000. |
Target
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Function
Tumor suppressor. Acts as a dual-specificity protein phosphatase, dephosphorylating tyrosine-, serine- and threonine-phosphorylated proteins. Also acts as a lipid phosphatase, removing the phosphate in the D3 position of the inositol ring from phosphatidylinositol 3,4,5-trisphosphate, phosphatidylinositol 3,4-diphosphate, phosphatidylinositol 3-phosphate and inositol 1,3,4,5-tetrakisphosphate with order of substrate preference in vitro PtdIns(3,4,5)P3 > PtdIns(3,4)P2 > PtdIns3P > Ins(1,3,4,5)P4. The lipid phosphatase activity is critical for its tumor suppressor function. Antagonizes the PI3K-AKT/PKB signaling pathway by dephosphorylating phosphoinositides and thereby modulating cell cycle progression and cell survival. The unphosphorylated form cooperates with AIP1 to suppress AKT1 activation. Dephosphorylates tyrosine-phosphorylated focal adhesion kinase and inhibits cell migration and integrin-mediated cell spreading and focal adhesion formation. Plays a role as a key modulator of the AKT-mTOR signaling pathway controlling the tempo of the process of newborn neurons integration during adult neurogenesis, including correct neuron positioning, dendritic development and synapse formation. May be a negative regulator of insulin signaling and glucose metabolism in adipose tissue. The nuclear monoubiquitinated form possesses greater apoptotic potential, whereas the cytoplasmic nonubiquitinated form induces less tumor suppressive ability. In motile cells, suppresses the formation of lateral pseudopods and thereby promotes cell polarization and directed movement.
Isoform alpha: Functional kinase, like isoform 1 it antagonizes the PI3K-AKT/PKB signaling pathway. Plays a role in mitochondrial energetic metabolism by promoting COX activity and ATP production, via collaboration with isoform 1 in increasing protein levels of PINK1. -
Tissue specificity
Expressed at a relatively high level in all adult tissues, including heart, brain, placenta, lung, liver, muscle, kidney and pancreas. -
Involvement in disease
Cowden syndrome 1
Lhermitte-Duclos disease
Bannayan-Riley-Ruvalcaba syndrome
Squamous cell carcinoma of the head and neck
Endometrial cancer
PTEN mutations are found in a subset of patients with Proteus syndrome, a genetically heterogeneous condition. The molecular diagnosis of PTEN mutation positive cases classifies Proteus syndrome patients as part of the PTEN hamartoma syndrome spectrum. As such, patients surviving the early years of Proteus syndrome are likely at a greater risk of developing malignancies.
Glioma 2
VACTERL association with hydrocephalus
Prostate cancer
Macrocephaly/autism syndrome
A microdeletion of chromosome 10q23 involving BMPR1A and PTEN is a cause of chromosome 10q23 deletion syndrome, which shows overlapping features of the following three disorders: Bannayan-Zonana syndrome, Cowden disease and juvenile polyposis syndrome. -
Sequence similarities
Contains 1 C2 tensin-type domain.
Contains 1 phosphatase tensin-type domain. -
Domain
The C2 domain binds phospholipid membranes in vitro in a Ca(2+)-independent manner; this binding is important for its tumor suppressor function. -
Post-translational
modificationsConstitutively phosphorylated by CK2 under normal conditions. Phosphorylated in vitro by MAST1, MAST2, MAST3 and STK11. Phosphorylation results in an inhibited activity towards PIP3. Phosphorylation can both inhibit or promote PDZ-binding. Phosphorylation at Tyr-336 by FRK/PTK5 protects this protein from ubiquitin-mediated degradation probably by inhibiting its binding to NEDD4. Phosphorylation by ROCK1 is essential for its stability and activity. Phosphorylation by PLK3 promotes its stability and prevents its degradation by the proteasome.
Monoubiquitinated; monoubiquitination is increased in presence of retinoic acid. Deubiquitinated by USP7; leading to its nuclear exclusion. Monoubiquitination of one of either Lys-13 and Lys-289 amino acid is sufficient to modulate PTEN compartmentalization. Ubiquitinated by XIAP/BIRC4. -
Cellular localization
Secreted. May be secreted via a classical signal peptide and reenter into cells with the help of a poly-Arg motif and Cytoplasm. Nucleus. Nucleus, PML body. Monoubiquitinated form is nuclear. Nonubiquitinated form is cytoplasmic. Colocalized with PML and USP7 in PML nuclear bodies. XIAP/BIRC4 promotes its nuclear localization. - Information by UniProt
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Database links
- Entrez Gene: 5728 Human
- Entrez Gene: 19211 Mouse
- Entrez Gene: 50557 Rat
- Omim: 601728 Human
- SwissProt: P60484 Human
- SwissProt: O08586 Mouse
- Unigene: 500466 Human
- Unigene: 729457 Human
see all -
Alternative names
- 10q23del antibody
- BZS antibody
- DEC antibody
see all
Images
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All lanes : Anti-PTEN (phospho T366) antibody [EP229] - BSA and Azide free (ab208104) at 1/100000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 100ng/ml Calyculin A for 30 minutes whole cell lysate
Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 100ng/ml Calyculin A for 30 minutes whole cell lysate, then the membrane was incubated with Alkaline phosphatase
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 47 kDa
Exposure time: 80 secondsThis data was developed using the same antibody clone in a different buffer formulation - BSA and Azide free (ab208104).
Recommended concentration for ab109454: 1/10000 dilution (0.1ug/ml).
Blocking and diluting buffer and concentration: 5% NFDM/TBST,
Primary and secondary antibodies were incubated for 1h at room temperature.
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All lanes : Anti-PTEN (phospho T366) antibody [EP229] - BSA and Azide free (ab208104) at 1/100000 dilution
Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) treated with 100 ng/ml Calyculin A for 30 minutes whole cell lysate
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) treated with 100 ng/ml Calyculin A for 30 minutes whole cell lysate, then the membrane was incubated with Alkaline phosphatase.
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 47 kDa
Exposure time: 40 secondsThis data was developed using the same antibody clone in a different buffer formulation - BSA and Azide free (ab208104).
Recommended concentration for ab109454: 1/10000 dilution (0.1ug/ml).
Blocking and diluting buffer and concentration: 5% NFDM/TBST,
Primary and secondary antibodies were incubated for 1h at room temperature.
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All lanes : Anti-PTEN (phospho T366) antibody [EP229] - BSA and Azide free (ab208104) at 1/100000 dilution
Lane 1 : C6 (Rat glial tumor glial cell) whole cell lysate
Lane 2 : C6 (Rat glial tumor glial cell) treated with 100 ng/ml Calyculin A for 30 minutes whole cell lysate
Lane 3 : C6 (Rat glial tumor glial cell) treated with 100 ng/ml Calyculin A for 30 minutes whole cell lysate, then the membrane was incubated with Alkaline phosphatase
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 47 kDa
Exposure time: 20 secondsThis data was developed using the same antibody clone in a different buffer formulation - BSA and Azide free (ab208104).
Recommended concentration for ab109454: 1/10000 dilution (0.1ug/ml).
Blocking and diluting buffer and concentration: 5% NFDM/TBST,
Primary and secondary antibodies were incubated for 1h at room temperature.
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This data was developed using the same antibody clone in a different buffer formulation - BSA and Azide free (ab208104).
Recommended concentration for ab109454: 1/200 dilution.
The human prostate cancer sections were performed by Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins, then treated with or without alkaline phosphatase. After alkaline phosphatase treatment, the sections were labelled with ab208104 at 1/2000 dilution for 30 mins at room temperature. The Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) were used as secondary antibody. The sections Counterstained with hematoxylin. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Human prostate cancer without alkaline phosphatase treatment (image A), no signal was detected when treated with alkaline phosphatase (image B).
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This data was developed using the same antibody clone in a different buffer formulation - BSA and Azide free (ab208104).
Recommended concentration for ab109454: 1/1000 dilution.
Dot blot analysis of PTEN (pT366) peptide (Lane 1), and PTEN non-phospho peptide (Lane 2) incubated with ab208104 at a dilution of 1/10000.
ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100,000.Exposure time: 180s
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (12)
ab109454 has been referenced in 12 publications.
- Xing Y et al. Methylated Vnn1 at promoter regions induces asthma occurrence via the PI3K/Akt/NF?B-mediated inflammation in IUGR mice. Biol Open 9:N/A (2020). PubMed: 32139393
- Li B et al. The MSC-Derived Exosomal lncRNA H19 Promotes Wound Healing in Diabetic Foot Ulcers by Upregulating PTEN via MicroRNA-152-3p. Mol Ther Nucleic Acids 19:814-826 (2020). PubMed: 31958697
- Engqvist H et al. Immunohistochemical validation of COL3A1, GPR158 and PITHD1 as prognostic biomarkers in early-stage ovarian carcinomas. BMC Cancer 19:928 (2019). PubMed: 31533654
- Stump B et al. Glycogen synthase kinase 3-ß inhibition induces lymphangiogenesis through ß-catenin-dependent and mTOR-independent pathways. PLoS One 14:e0213831 (2019). PubMed: 30964887
- Gong L et al. miR-222 promotes invasion and migration of ovarian carcinoma by targeting PTEN. Oncol Lett 16:984-990 (2018). PubMed: 29963173