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. Specific for phospho mTOR, giving a band at approx 250 kD.
Note: Not all species have been tested for usefulness with this product. Only those species listed have been tested. We cannot make any guarantees about additional reactivities which may or may not occur.
This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to amino acids 2440-2457 of human mTOR.
Localization:
Cell Membrane and Cytoplasmic
Modification:
p Ser2448
Species Reactivity:
Cross-reacts with Human. Not yet tested in other species.
Applications:
Uses:
This antibody is useful in ELISA, Western Blot and Immunohistochemistry.
Dilutions:
ELISA,
Immunohistochemistry,
Immunohistochemistry-Paraffin 1/50-1/100,
Western Blot 1/500-1/1000
Unit Size:
0.1 mg
Concentration:
1.5 mg/ml
Packaging:
Storage:
Aliquot and store at -20 °C or -80 °C. Avoid freeze-thaw cycles.
Buffer:
0.02 M Potassium Phosphate, with 0.15 M Sodium Chloride [pH 7.2]
Preservative:
0.01% Sodium Azide
Limitations:
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Products are guaranteed for 6 months from date of receipt, except for peptides and proteins which are guaranteed for 3 months.
Mammalian target of rapamycin (mTOR) is a serine and threonine protein kinase that regulates numerous cellular functions, in particular, the initiation of protein translation. Rapamycin is a natural product macrolide that induces G1 growth arrest in yeast, Drosophila, and mammalian cells. mTOR is one of a family of proteins involved in cell cycle progression, DNA recombination, and DNA damage detection. In rat, mTOR is a 245-kD protein referred to as RAFT1 with significant homology to the S. cerevisiae protein TOR1 and has been shown to associate with the immunophilin FKBP12 in a rapamycin-dependent fashion. The FKBP12-rapamycin complex is known to inhibit progression through the G1 cell cycle stage by interfering with mitogenic signaling pathways involved in G1 progression in several cell types, as well as in yeast. The binding of mTOR to FKBP12-rapamycin correlates with the ability of these ligands to inhibit cell cycle progression.
Western Blot: mTOR [p Ser2448] Antibody [NB600-607] - Figure 1. Immunoblotting. Affinity Purified Anti-mTOR pS2448 (Rabbit) is shown to detect a 250 kDa band (indicated) corresponding to phosphorylated human mTOR present in a 293T whole cell lysates. Cells were serumstarved for 24 hours prior to harvest. ~20ìg 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.
Immunohistochemistry: mTOR [p Ser2448] Antibody [NB600-607] - Immunohistochemistry. Affinity purified anti-mTOR pS2448 antibody was used at 5 ìg/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. 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.