Anti mTOR, phospho Ser2448  

Rockland's affinity purified anti-mTOR pS 2448 antibody was used at 5 μg/ml to detect signal in a variety of tissues including multi-human, multi-brain and multi-cancer slides. This image shows moderate staining of proximal convoluted tubules of the kidney (40X). Tissue was formalin-fixed and paraffin embedded. The image shows localization of the antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain. Personal Communication, Tina Roush, LifeSpanBiosciences, Seattle, WA.

Affinity Purified Anti-mTOR pS 2448 (Rabbit) is shown to detect a 250 kDa band (indicated) corresponding to phosphorylated human mTOR present in a 293T whole cell lysates. Cells were serum-starved for 24 hours prior to harvest. ~20 ug of lysate was loaded per lane for SDS-PAGE. Untreated cells are shown in lane 1, whereas cells in lane 2 were treated with IGF-1 (100 ng/ml) for 20 min prior to harvest. Follow reaction of antibody with a 1:2000 dilution of HRP Goat-a-Rabbit IgG for visualization.

ELISA Results of Rabbit Anti-mTOR pS2448 Antibody tested against BSA-conjugated non-phospho [purple line] and phospho [blue line] forms of immunizing peptide. Each well was coated in duplicate with either 0.1μg of conjugate. The working dilution is 1:37,000. The starting dilution of antibody was 5μg/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using HRP conjugate stabilizer (p/n MB-067), Goat Anti-Rabbit HRP conjugated (p/n 611-103-122) and TMB substrate (p/n TMBE-1000).

Metabolic and autocrine regulation of the mTOR pathway by Beta-cells.

Effect of exercise and insulin stimulation with NAC or CON infusion on phosphorylation of protein translation regulation proteins mTOR Ser2448？(A), p70S6K Thr389？(B), S6 Ser235/236？(C), eukaryotic initiation factor 4E-binding protein 1 (4E-BP1) Thr37/46？(D), and STAT3 Tyr705？(E). Western blots normalized to Ponceau S stain for each lane, and representative blots from 1 participant are shown. Data are means ± SD for？n？= 7. *Significantly different, NAC vs. CON (P？< 0.05); †significantly different from BASE (P？< 0.05). Fig 6. PMID:？26105008

Protein synthesis in differentiated myotubes. (A) Phenylanine incorporation into protein in myotubes from spinal cord‐injured (gray bars) and able‐bodied (white bars) individuals. Bars represent mean？±？SD and individual data points are overlaid. (B) Phosphorylated and total protein content of molecules in the Akt‐mTOR signalling axis (pAkt(Ser473), pAkt(Thr308), total Akt, mTOR？(Ser2448), total mTOR, Raptor, p4EBP1(Ser37/46), total 4EBP, pS6(Ser235/236), total S6) in myotubes from spinal cord‐injured (gray bars) and able‐bodied (white bars) individuals. Bars represent mean？±？SD and individual data points are overlaid.？n？=？6？8; Mann？Whitney test (significance？P？<？0.05). (C) Representative Western blot images. mTOR, mechanistic target of rapamycin; 4EBP, 4E binding protein. Fig 3. PMID:？29906337

Immunolocalization and fluorescence intensity of pmTOR. (A) Representative confocal images showing the organ of Corti immunolabelled with anti-pmTOR (green) and counterstained with bisbenzimide nuclear dye (blue) of all experimental groups and for each cochlear location (from B1 to H). Scale bar: 50 μm. L: lateral, M: medial. (B) Fluorescence intensity analysis of pmTOR immunostaining. The graph shows the signal intensity of each sample for all cochlear locations (black: NH, red: untreated, green: BDNF-treated). The same symbol was used to identify the BDNF-treated and untreated ears of individual animals. Fig 8.PMID:？36428503

Protein quantification of pmTOR (p/n 600-401-422) and mTOR (p/n 600-401-897). (A) Representative Western blot bands (original Western blot bands are shown in？Supplementary Figure S2). Western blot analysis of pmTOR/mTOR (B), mTOR/GAPDH (C) and pmTOR/GAPDH (D) on organ of Corti samples from all the experimental groups. Histograms show mean ± SD; the dots indicate the densitometric values for individual samples (n？= 5; each sample is a pool of two organs of Corti). Fig 11.PMID:？36428503