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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|
|Mouse, Rat, Human
||Lot dependent; please refer to CoA along with shipment
|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
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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 TA300537.
Immunocytochemistry/ Immunofluorescence-mTOR antibody [Y391](TA300537); ICC/IF image of TA300537 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 1.43uM.
Flow Cytometry-mTOR antibody [Y391] - ChIP Grade(TA300537); Overlay histogram showing HeLa cells stained with TA300537 (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 (1ug/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.