Phospho-mTOR (S2448) Mouse Monoclonal Antibody [A5D5]
cat.: HA600094
Product Type: Mouse monoclonal IgG1, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P
Clonality: Monoclonal
Clone number: A5D5
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage buffer: PBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 2ug/ul
Purification: Protein G affinity purified.
Molecular weight: Predicted band size: 289 kDa
Isotype: IgG1
Immunogen: Synthetic peptide within human mTOR aa 2430-2480.
Positive control: HeLa cell lysate, HeLa starved overnight then treated with 200nM PMA for 4 hours cell lysate, NIH/3T3 cell lysate, NIH/3T3 starved for 16 hours then treated with 100nM insulin for 10 minutes cell lysate, human liver tissue.
Subcellular location: Lysosome, Mitochondrion outer membrane, Endoplasmic reticulum membrane, Cytoplasm.
Recommended Dilutions:
  WB
  IHC-P
1:1,000-1:2,000
1:50-1:200
Uniprot #: SwissProt: P42345 Human | Q9JLN9 Mouse
Alternative names: dJ576K7.1 (FK506 binding protein 12 rapamycin associated protein 1) FK506 binding protein 12 rapamycin associated protein 1 FK506 binding protein 12 rapamycin associated protein 2 FK506 binding protein 12 rapamycin complex associated protein 1 FK506-binding protein 12-rapamycin complex-associated protein 1 FKBP rapamycin associated protein FKBP12 rapamycin complex associated protein FKBP12-rapamycin complex-associated protein 1 FKBP12-rapamycin complex-associated protein FLJ44809 FRAP FRAP1 FRAP2 Mammalian target of rapamycin Mechanistic target of rapamycin mTOR MTOR_HUMAN OTTHUMP00000001983 RAFT1 Rapamycin and FKBP12 target 1 Rapamycin associated protein FRAP2 Rapamycin target protein 1 Rapamycin target protein RAPT1 Serine/threonine-protein kinase mTOR
Images
HA600094_1.jpg Fig1: Western blot analysis of Phospho-mTOR (S2448) on different lysates with Mouse anti-Phospho-mTOR (S2448) antibody (HA600094) at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HeLa starved overnight then treated with 200nM PMA for 4 hours cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 289 kDa
Observed band size: 289 kDa

Exposure time: 1 minute 30 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA600094) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
HA600094_2.jpg Fig2: Western blot analysis of Phospho-mTOR (S2448) on different lysates with Rabbit anti-Phospho-mTOR (S2448) antibody (HA600094) at 1/1,000 dilution.

Lane 1: NIH/3T3 (Mouse fibroblasts) cell lysate
Lane 2: NIH/3T3 starved for 16 hours then treated with 100nM insulin for 10 minutes cell lysate

Lysates/proteins at 10 µg/Lane.
Exposure time: 180 seconds; ECL: K1802

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA600094, 1/1,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Mouse IgG-HRP (HA1006), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 288.9 kDa
Observed band size: 289 kDa
HA600094_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human liver tissue with Mouse anti-Phospho-mTOR (S2448) antibody (HA600094) at 1/100 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA600094) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.