Human Phospho-Akt1 (T308) Antibody

Detection of Human and Mouse Phospho-Akt1 (T308) by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line and NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human PDGF-BB (Catalog # 220-BB) for 20 minutes or 100nM Calyculin A for 30 minutes. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Phospho-Akt1 (T308) Monoclonal Antibody (Catalog # MAB7419) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Phospho-Akt1 (T308) at approximately 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Phospho-Akt1 (T308) in CCD‑1070Sk Human Cell Line. Akt1 phosphorylated at T308 was detected in immersion fixed CCD-1070Sk human foreskin fibroblast cell line stimulated with Recombinant Human PDGF-BB (Catalog # 220-BB) using Mouse Anti-Human Phospho-Akt1 (T308) Mono-clonal Antibody (Catalog # MAB7419) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the Northern-Lights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human AKT1 by Western Blot ASPP2 blocks Gal-1 dependent nanoclustering and halts oncogenic H-ras induced transformation.Nanoclustering-FRET analysis in HEK cells coexpressing mGFP- and mCherry-tagged (A, B) H-rasG12V or (C, D) K-rasG12V. Cells were analysed after overexpression of either Gal-1 or ASPP2 plasmids, or both (1:1 ratio). Plotted are the means ± SEM, n = 3. (E) Representative Western blots from HEK cells expressing mGFP-H-rasG12V (left) or K-rasG12V (right) alone or together with Gal-1 and ASPP2. Statistical significance of differences was examined using t-test (n = 3; *, p<0.05, **, p< 0.01, ***, p<0.001). (F, G) Colony survival assay of NIH/3T3 cells stably expressing (F) H-rasG12V or (G) K-rasG12V and transiently expressing indicated constructs. Colony survival was graphed based on mean foci areas calculated from at least 4 independent biological repeats. (A-D, F-G) Statistical significance of differences between controls and treated samples was examined using one-way ANOVA (ns, not significant; *, p<0.05; **, p<0.01; ****, p<0.0001). Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0159677), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human AKT1 by Western Blot ASPP2 increases oncogenic H-ras, K-ras and N-ras-effector-recruitment, as well as ERK- and AKT-signalling.(A) Left, scheme explaining effector-recruitment FRET analysis in HEK cells. Right, examples of FLIM-FRET images of HEK cells from the different FRET samples as indicated. (B-D) Effector-recruitment FRET analysis in HEK cells coexpressing (B) mGFP-H-rasG12V, (C) mGFP-K-rasG12V or (D) mGFP-NrasG12V and mRFP-RBD from C-Raf. The effect of Gal-1 or ASPP2 expression on effector-recruitment FRET was compared to control samples. (E) Representative Western blots from HEK cells expressing mGFP-H-rasG12V (left), K-rasG12V (middle) or N-rasG12V (right) without or with Gal-1 or ASPP2. Statistical significance of differences between controls and treated samples was examined using one-way ANOVA (mean ± SEM, n = 3; ns, not significant; *, p<0.05, **, p< 0.01, ***, p<0.001, ****, p< 0.0001). Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0159677), licensed under a CC-BY license. Not internally tested by R&D Systems.