Detection of Human/Mouse/Rat PTEN by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and rat and mouse brain tissue. PVDF membrane was probed with 0.5 µg/mL Mouse Anti-Human/Mouse/Rat PTEN Monoclonal Antibody (Catalog # MAB847) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). For additional reference, Recombinant Human PTEN (Catalog # 847‑PN) (5 ng) was included. A specific band for PTEN was detected at approximately 54 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4.
Detection of PTEN in Human PBMC lymphocytes by Flow Cytometry.
Human peripheral blood mononuclear cell lymphocytes were stained with Mouse Anti-Human/Mouse/Rat PTEN Monoclonal Antibody (Catalog # MAB847, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')2 Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.
PTEN in Human Liver.
PTEN was detected in immersion fixed paraffin-embedded sections of human liver array using Mouse Anti-Human/Mouse/Rat 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.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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.
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).
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.
Phosphatase and Tensin Homolog Deleted on Chromosome 10
Entrez Gene IDs:
5728 (Human); 19211 (Mouse); 50557 (Rat)
BZS; EC 22.214.171.124,10q23del; EC 126.96.36.199; EC 188.8.131.52; GLM2; MMAC1 phosphatase and tensin homolog deleted on chromosome 10; MMAC1; MMAC1MGC11227; Mutated in multiple advanced cancers 1; phosphatase and tensin homologDEC; phosphatidylinositol-34,5-trisphosphate 3-phosphatase and dual-specificityprotein phosphatase PTEN; PTEN; PTEN1; TEP1MHAM
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