mTOR antibody

• Catalog Number: orb345523
• Category: Antibodies
• Description: mTOR antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, WB
• Reactivity: Human
• Isotype: IgG
• Immunogen: This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to an internal region near amino acids 2430-2460 of human mTOR.
• Concentration: 0.9 mg/ml
• Dilution range: ELISA: 1:50,000 - 1:100,000, WB: 1:250 - 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 both phosphorylated and non-phosphorylated forms of mTOR at S2448 from human derived tissues and cells. A BLAST analysis was used to suggest cross reactivity with mTOR protein from rat and mouse based on 100% homology with the immunizing sequence. Expect partial reactivity against mTOR homologues from zebrafish (94%) and dog (89%). Reactivity against homologues from other sources is not known.
• 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 mTOR antibody has been tested for use in ELISA and western blotting. ELISA data demonstrate reactivity against both phosphorylated and non-phosphorylated mTOR at S2448 and western blotting shows a band at approximately 250 kDa. Reactivity in other immunoassays is unknown.
• Expiration Date: 12 months from date of receipt.

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

ELISA results of purified Rabbit anti-mTOR Antibody tested against BSA-conjugated peptide of immunizing peptide. Each well was coated in duplicate with 0.1 µg of conjugate. 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 3% fish gel, Goat anti-Rabbit IgG Antibody Peroxidase Conjugated (Min X Bv Ch Gt GP Ham Hs Hu Ms Rt & Sh Serum Proteins) (p/n orb347654) and TMB ELISA Peroxidase Substrate (p/n orb348651).

Western blot using Biorbyt's Affinity Purified anti-mTOR antibody shows detection of a band ~245 kDa corresponding to human mTor (arrowhead). Approximately 30 µg of HEK293 cell lysate (p/n orb348669) was separated by 4-8% SDS-PAGE and transferred onto nitrocellulose. After blocking, the membrane was probed with the primary antibody diluted to 1:650 for 2h at RT. The membrane was washed and reacted with a 1:10000 dilution of IRDye™800 conjugated Gt-a-Rabbit IgG [H&L] MX for 45 min at room temperature.