mTOR (phospho-S2448) antibody

• Catalog Number: orb345415
• Category: Antibodies
• Description: mTOR (phospho-S2448) antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IHC, WB
• Reactivity: Human
• Isotype: IgG
• Immunogen: Anti-mTOR pS2448 antibody was prepared from whole rabbit serum produced by repeated immunizations with a phosphorylated synthetic peptide corresponding a c-terminal region near Serine 2448 of human mTOR.
• Concentration: 1.1mg/mL
• Dilution range: ELISA: 1:10,000 - 1:100,000, IHC: 5.0 µg/ml, WB: 1:500 - 1:2,000
• Form/Appearance: Liquid (sterile filtered)
• Purity: This is an affinity purified antibody produced by immunoaffinity chromatography using the immunizing peptide after immobilization to a solid phase. Reactivity occurs with phosphorylated mTOR from human derived tissues and cells. Reactivity against mTOR from other species has not been determined, however, reactivity with mouse and rat is suggested based on protein sequence homologies.
• Conjugation: Unconjugated
• UniProt ID: P42345
• NCBI: 1169735
• Storage: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Alternative names: rabbit anti-mTOR pS2448 antibody, FKBP12 rapamycin
• Note: For research use only
• Application notes: This affinity purified antibody has been tested for use in immunohistochemistry, ELISA and western blotting. Western blotting shows reactivity specific for phospho mTOR detecting a band at approximately 250 kDa. Reactivity in other immunoassays is unknown.
• Expiration Date: 12 months from date of receipt.

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.

Biorbyt'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.

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 0.1 µg of conjugate. The working dilution is 1:37000. The starting dilution of antibody was 5 µg/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, Goat Anti-Rabbit HRP conjugated (p/n orb347654) and TMB substrate (p/n orb348651).

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