Western blot shows lysates of mouse brain tissue. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human/Mouse PTEN Monoclonal Antibody (Catalog # MAB847) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for PTEN at approximately 54 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.**For Western blot, we recommend the use of Rabbit Anti-Human/Mouse/Rat PTEN Affinitiy-purified Polyclonal Ab (AF847).

PTEN in Human Liver.

PTEN was detected in immersion fixed paraffin-embedded sections of human liver array using Mouse Anti-Human/Mouse PTEN Monoclonal Antibody (Catalog # MAB847) at 25 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of PTEN in Human U937 cell line by Flow Cytometry.

Human U937 histiocytic lymphoma cell line was stained with Mouse Anti-Human/Mouse PTEN Monoclonal Antibody (Catalog # MAB847, filled histogram) or isotype control antibody (MAB002, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG F(ab')2Secondary Antibody (F0101B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (FC005). Staining was performed using our protocol for Staining Intracellular Molecules.

Detection of PTEN in Human PBMC lymphocytes by Flow Cytometry.

Human peripheral blood mononuclear cell lymphocytes were stained with Mouse Anti-Human/Mouse PTEN Monoclonal Antibody (Catalog # MAB847, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')₂Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Western Blot Shows Human PTEN Specificity by Using Knockout Cell Line.

Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and PTEN knockout HeLa cell line (KO). PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human/Mouse PTEN Monoclonal Antibody (Catalog # MAB847) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for PTEN at approximately 55 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Detection of Rat PTEN by Western Blot

Western blot analysis of apoptotic-related proteins after furin siRNA transfection.Gramulosa cells were transfected with furin siRNA or control siRNA for 48 h before being subjected to protein extraction and Western blot with the indicated antibodies. Shown at the left were representative Western blot images. Three such experiments were quantified by measuring the intensity of apoptotic-related proteins relative to the alpha-tubulin (loading) control. The anti-apoptotic proteins XIAP and p-Akt were decreased by appropriate 3-folds (**, P<0.01) and 2-folds (**, P<0.01). On the contrary, the levels of pro-apoptotic, cleaved caspase-3 and PTEN were increased appropriate 4-folds (**, P<0.01) and 4.5-folds (**, P<0.01). Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0050479), licensed under a CC-BY license. Not internally tested by R&D Systems.

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Applications for Human/Mouse PTEN Antibody

Application

Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunohistochemistry

8-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human liver

Intracellular Staining by Flow Cytometry

0.25 µg/10⁶ cells
Sample: Human peripheral blood lymphocytes and U937 cell line fixed with paraformaldehyde and permeabilized with saponin

Knockout Validated

PTEN is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in PTEN knockout HeLa cell line.

Western Blot

Mouse brain tissue.
**For Western blot, we recommend Rabbit Anti-Human/Mouse/Rat PTEN Affinity-purified Polyclonal Ab (Catalog # AF847)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: PTEN

The tumor suppressor gene PTEN (phosphatase and tensin homolog deleted on chromosome 10), also known as MMAC1 (mutated in multiple advanced cancers 1), encodes a phosphatase that contains the catalytic signature motif (HCxxGxxRS/T) found in all members of the protein tyrosine phosphatase family. In vitro, the recombinant PTEN has both lipid phosphatase and protein phosphatase activities (1, 2). Interestingly, accumulating evidence has shown that the tumor suppressor activity of PTEN relies on its ability to dephosphorylate phosphatidylinositol (3,4,5)-triphosphate specifically at position 3 of the inositol ring (3). This activity reduces the levels of phosphatidylinositol (3,4,5)-triphosphate which is specifically produced from phosphatidylinositol (4,5)-diphosphate by PI 3-kinase upon activation by a variety of stimuli. Therefore, PTEN antagonizes PI 3-kinase-induced downstream signaling events and cellular processes including cell growth, apoptosis and cell motility. In vivo, the importance of PTEN catalytic activity in its tumor suppressor functions is underscored by the fact that the majority of PTEN missense mutations detected in tumor specimens target the phosphatase domain and cause a loss in PTEN phosphatase activity (4).

References

Maehama, T. and J. Dixon (1998) J. Biol. Chem. 273:13375.
Das, S. et al. (2003) Proc. Natl. Acad. Sci. USA 100:7491.
Myers, M. et al. (1998) Proc. Natl. Acad. Sci. USA 95:13513.
Waite, K. and C. Eng (2002) Am. J. Hum. Genet. 70:829.

Long Name

Phosphatase and Tensin Homolog Deleted on Chromosome 10

Alternate Names

MMAC1

Entrez Gene IDs

5728 (Human); 19211 (Mouse); 50557 (Rat)

Gene Symbol

PTEN

UniProt

P60484 (Human)

Additional PTEN Products

Product Documents for Human/Mouse PTEN Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human/Mouse PTEN Antibody

This product is covered by the following U.S. patent: USSN # 10/299,003.

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human/Mouse PTEN Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB847

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Applications