Human/Mouse PTEN Antibody

Detection of Mouse PTEN by Western Blot. 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.