Recombinant Anti-PTEN antibody [EPR22636-122] KO Tested (ab267787)

Product name

Anti-PTEN antibody [EPR22636-122]
See all PTEN primary antibodies
Description

Rabbit monoclonal [EPR22636-122] to PTEN
Host species

Rabbit
Tested applications

Suitable for: WB, IHC-P, IP, Flow Cyt (Intra)more details
Unsuitable for: ICC/IF
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Wild-type HAP1, MCF7, MDA-MB-468, HeLa, C6, RAW264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse kidney, Mouse spleen and Rat lung lysates. IHC-P: Human endometrial cancer, Human ovarian cancer, Human pancreas, Mouse pancreas and Rat pancreas tissues. Flow Cyt (intra): HeLa and NIH/3T3 cells. IP: MCF7 and HeLa cells.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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, 0.05% BSA, 40% Glycerol (glycerin, glycerine)
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR22636-122
Isotype

IgG
Research areas
- Neuroscience
- Development

Alternative Versions
- Anti-PTEN antibody [EPR22636-122] - BSA and Azide free (ab267791)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab267787 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
WB		1/1000. Detects a band of approximately 54 kDa (predicted molecular weight: 47 kDa).
IHC-P		1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP		1/30.
Flow Cyt (Intra)		1/500.

Notes
WB 1/1000. Detects a band of approximately 54 kDa (predicted molecular weight: 47 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP 1/30.
Flow Cyt (Intra) 1/500.

Application notes

Is unsuitable for ICC/IF.

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
modifications

Constitutively 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.
Target information above from: UniProt accession P60484 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
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
- Unigene: 245395 Mouse
see all
Alternative names
- 10q23del antibody
- BZS antibody
- DEC antibody
- GLM2 antibody
- MGC11227 antibody
- MHAM antibody
- MMAC1 antibody
- MMAC1 phosphatase and tensin homolog deleted on chromosome 10 antibody
- Mutated in multiple advanced cancers 1 antibody
- Phosphatase and tensin homolog antibody
- Phosphatase and tensin like protein antibody
- Phosphatidylinositol 3,4,5-trisphosphate 3-phosphatase and dual-specificity protein phosphatase PTEN antibody
- Pten antibody
- PTEN_HUMAN antibody
- PTEN1 antibody
- TEP1 antibody
see all

Western blot - Anti-PTEN antibody [EPR22636-122] (ab267787)

All lanes : Anti-PTEN antibody [EPR22636-122] (ab267787) at 1/1000 dilution

Lane 1 : C6 (rat glial tumor glial cell), whole cell lysate
Lane 2 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
Lane 3 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate
Lane 4 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate
Lane 5 : Mouse kidney tissue lysate
Lane 6 : Mouse spleen tissue lysate
Lane 7 : Rat lung tissue lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 47 kDa
Observed band size: 54 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure times:

Lanes 1-6: 3 seconds;
Lane 7: 7 seconds.

The molecular weight observed is consistent with what has been described in the literature (PMID:10514407).
Western blot - Anti-PTEN antibody [EPR22636-122] (ab267787)

All lanes : Anti-PTEN antibody [EPR22636-122] (ab267787) at 1/1000 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : PTEN knockout HAP1 whole cell lysate
Lane 3 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate
Lane 4 : MDA-MB-468 (human breast adenocarcinoma epithelial cell), whole cell lysate
Lane 5 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 47 kDa
Observed band size: 54 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure times:

Lanes 1-4: 3 seconds;
Lane 5: 114 seconds.

ab267787 was shown to specifically react with PTEN in wild-type HAP1 cells as signal was lost in PTEN knockout cells. Wild-type and PTEN knockout samples were subjected to SDS-PAGE. Ab267787 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/20,000 dilution for 1 hour at room temperature before imaging.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:10514407).

Negative control: MDA-MB-468 (PMID:21358673,15674339).
Immunoprecipitation - Anti-PTEN antibody [EPR22636-122] (ab267787)

PTEN was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate with ab267787 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab267787 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug

Lane 2: ab267787 IP in HeLa whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab267787 in HeLa whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 15 seconds
Immunoprecipitation - Anti-PTEN antibody [EPR22636-122] (ab267787)

PTEN was immunoprecipitated from 0.35 mg MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate with ab267787 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab267787 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1: MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10ug

Lane 2: ab267787 IP in MCF7 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab267787 in MCF7 whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 15 seconds
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [EPR22636-122] (ab267787)

Immunohistochemical analysis of paraffin-embedded Rat pancreas tissue labeling PTEN with ab267787 at 1/2000 dilution (2.18 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat pancreas (PMID:11021813). The section was incubated with ab267787 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [EPR22636-122] (ab267787)

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling PTEN with ab267787 at 1/2000 dilution (2.18 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse pancreas (PMID:11021813). The section was incubated with ab267787 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [EPR22636-122] (ab267787)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling PTEN with ab267787 at 1/2000 dilution (2.18 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human pancreas (PMID:11021813). The section was incubated with ab267787 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [EPR22636-122] (ab267787)

Immunohistochemical analysis of paraffin-embedded Human ovarian cancer tissue labeling PTEN with ab267787 at 1/2000 dilution (2.18 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on stroma of human ovarian cancer (PMID:25608477). The section was incubated with ab267787 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [EPR22636-122] (ab267787)

Immunohistochemical analysis of paraffin-embedded Human endometrial cancer tissue labeling PTEN with ab267787 at 1/2000 dilution (2.18 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on stroma of human endometrial cancer (PMID:2230170). The section was incubated with ab267787 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Flow Cytometry (Intracellular) - Anti-PTEN antibody [EPR22636-122] (ab267787)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling PTEN with ab267787 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Flow Cytometry (Intracellular) - Anti-PTEN antibody [EPR22636-122] (ab267787)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling PTEN with ab267787 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Anti-PTEN antibody [EPR22636-122] (ab267787)

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.

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab267787? Please let us know so that we can cite the reference in this datasheet.

ab267787 has been referenced in 12 publications.

Wu Z et al. The inhibitory effect of human DEFA5 in growth of gastric cancer by targeting BMI1. Cancer Sci 112:1075-1083 (2021). PubMed: 33503272
Zhou X et al. Silencing of MEG3 attenuated the role of lipopolysaccharides by modulating the miR-93-5p/PTEN pathway in Leydig cells. Reprod Biol Endocrinol 19:33 (2021). PubMed: 33639974
Cheng Z et al. Upregulation of circRNA_100395 sponges miR-142-3p to inhibit gastric cancer progression by targeting the PI3K/AKT axis. Oncol Lett 21:419 (2021). PubMed: 33841580
Zhan D et al. Hypoxia-inducible factor-1a regulates PI3K/AKT signaling through microRNA-32-5p/PTEN and affects nucleus pulposus cell proliferation and apoptosis. Exp Ther Med 21:646 (2021). PubMed: 33968177
Yang H et al. LncRNA JPX regulates proliferation and apoptosis of nucleus pulposus cells by targeting the miR-18a-5p/HIF-1a/Hippo-YAP pathway. Biochem Biophys Res Commun 566:16-23 (2021). PubMed: 34111667

View all Publications for this product

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Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (12)

Customer reviews and Q&As

Post-translational
modifications