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Anti-FRAP1 Antibody Y391
Also for FRAP1 (NM_004958)
|A synthetic peptide corresponding to residues near the C-term of PI3K/PI4K domain of human mTOR/FRAP was used as an immunogen|
|Human, Mouse, Rat
||0.5~1.0 mg/ml (Lot Dependent)
|WB, IHC, IF, FC
||IHC-P: Use at an assay dependent dilution; WB: 1:1000 - 1:2000; ICC/IF: 1:50 - 1:100; FC: 1:50 - 1:80; IP: 1:50
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Tissue culture supernatant (Protein A or G Sepharose)
|Homo sapiens mechanistic target of rapamycin (serine/threonine kinase) (MTOR)|
|FRAP; FRAP1; FRAP2; RAFT1; RAPT1|
|The mammalian target of rapamycin (mTOR) is a major effector of cell growth and proliferation via the regulation of protein synthesis. mTOR is composed of an N-term; 20 tandem repeats- HEAT which are implicated in protein-protein interactions and a C-term; which includes a FAT domain, a FBR domain, a kinase domain, a NDR domain and a FATC domain. The FATC domain is essential to mTOR activity and the deletion of a single amino acid from this domain abrogates the activity. mTOR can be auto-phosphorylated via its intrinsic serine/threonine kinase activity. mTOR regulates protein synthesis through the phosphorylation and inactivation of the repressor of mRNA translation 4E-BP1 and though the phosphorylation and activation of S6 kinase. RAPTOR (regulatory associated protein of TOR) is a positive regulator of TOR. Other known mediators of mTOR include PI3-K and ATK from the insulin pathway. |
Senescence and Autophagy
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Western blot - mTOR antibody [Y391]; All lanes : Anti-mTOR antibody [Y391] - ChIP Grade at 1/2000 dilution.Lane 1 : Hela cell lysate.Lane 2 : MCF-7.Predicted band size : 289 kDa.Observed band size : >250 kDa .
Immunohistochemistry (Paraffin-embedded sections) - mTOR antibody [Y391]; Immunohistochemical analysis of mTOR expression in paraffin-embedded human prostate carcinoma, using 1/250 ab32028.
Immunocytochemistry/ Immunofluorescence-mTOR antibody [Y391](ab32028); ICC/IF image of ab32028 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody overnight at +4°C. The secondary antibody (green) was Alexa Fluor? 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor? 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Flow Cytometry-mTOR antibody [Y391] - ChIP Grade(ab32028); Overlay histogram showing HeLa cells stained with ab32028 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody for 30 min at 22°C. The secondary antibody used was DyLight? 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x10^6 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a significantly decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween used under the same conditions.