Human/Mouse/Rat PTEN Antibody

Detection of Human/Mouse/Rat PTEN by Western Blot. Western blot shows lysates of mouse and rat brain tissue, A431 human epithelial carcinoma cell line, and MRC-5 human embryonic lung fibroblast cell line. PVDF membrane was probed with 0.1 µg/mL Rabbit Anti-Human/Mouse/Rat PTEN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF847) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). For additional reference, recombinant human PTEN (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.

PTEN in Human Pancreas. PTEN was detected in immersion fixed paraffin-embedded sections of human pancreas using Rabbit Anti-Human/Mouse/Rat PTEN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF847) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in islet cells. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

PTEN in Mouse Pancreas. PTEN was detected in immersion fixed paraffin-embedded sections of mouse pancreas using Rabbit Anti-Human/Mouse/Rat PTEN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF847) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in islet cells. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of PTEN in Human PBMC lymphocytes by Flow Cytometry. Human peripheral blood lymphocytes were stained with Rabbit Anti-Human/Mouse/Rat PTEN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF847, filled histogram) or control antibody (Catalog # AB-105-C, open histogram), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Detection of Human PTEN by Simple Western^TM. Simple Western lane view shows lysates of A431 human epithelial carcinoma cell line and HeLa human cervical epithelial carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for PTEN at approximately 60 kDa (as indicated) using 1 µg/mL of Rabbit Anti-Human/Mouse/Rat PTEN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF847). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

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 0.1 µg/mL of Rabbit Anti-Human/Mouse/Rat PTEN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF847) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). 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 (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human PTEN by Western Blot FoxM1 inhibitor induced RASSF1A and PTEN expression and YAP phosphorylation in mCRC cells. T84 and Colo 205 cells were treated with FoxM1 inhibitor, thiostrepton (Th) with different doses (0–8 µM) for 48 h. (A,B) mRNA was extracted after 24 h for detection of RASSF1A by qRT-PCR. Bar graphs show quantitative results normalized to GAPDH mRNA levels. Cell lysates were monitored by immunoblot for FoxM1(C), RASSF1A (D), p-YAP and total YAP (E), PTEN (F), and GAPDH as indicated. Immunoblots were quantified by scanning densitometry and normalized against GAPDH expression (lower panels of (C–E) for FoxM1, RASSF1A and p-YAP, respectively. Right panel of (F) represents the quantification of PTEN). (G) Patient derived organoids were treated with (8 µM) and without thiostrepton for 24 h and 48 h. Organoid cell lysates were analyzed by immunoblot for RASSF1A expression and quantified by densitometry ((G), lower panel). The results are from three independent experiments. (* p < 0.05, ** p < 0.01, *** p < 0.001). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30744076), licensed under a CC-BY license. Not internally tested by R&D Systems.