Anti-Phospho-mTOR (Ser2481) Polyclonal Antibody is a Rabbit antibody targeting Phospho-mTOR (Ser2481). Anti-Phospho-mTOR (Ser2481) Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Dog,Pig,Horse,Rabbit,Sheep)

1. Blank control: A431. Primary Antibody (green line): Rabbit Anti-Phospho-mTOR (Ser2481) antibody (TMAB-01459)
Dilution: 1 μg/Test;
Secondary Antibody: Goat anti-rabbit IgG-AF488
Dilution: 0.5 μg/Test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
2. A431 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (Phospho-mTOR (Ser2481)) polyclonal Antibody, Unconjugated (TMAB-01459) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
3. Sample:
Hepg2 (Human) Cell Lysate at 30 μg
Primary: Anti-Phospho-mTOR (Ser2481) (TMAB-01459) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 289 kDa
Observed band size: 289 kDa
4. Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-Phospho-mTOR (Ser2481) Polyclonal Antibody, Unconjugated (TMAB-01459) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
5. Paraformaldehyde-fixed, paraffin embedded (mouse brain tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Phospho-mTOR (Ser2481)) Polyclonal Antibody, Unconjugated (TMAB-01459) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
6. Blank control: mouse splenocytes (blue)
Isotype Control Antibody: Rabbit Igg (orange);
Secondary Antibody: Goat anti-rabbit IgG-FITC (white blue),
Dilution: 1:100 in 1 X PBS containing 0.5% BSA;
Primary Antibody Dilution: 5 μL in 100 μL 1X PBS containing 0.5% BSA (green).
7. Blank control (blue line): Hela (fixed with 70% ethanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 30 min on ice)
Primary Antibody (green line): Rabbit Anti-Phospho-mTOR (Ser2481) antibody (TMAB-01459), Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1 μg/test.
8. Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Phospho-mTOR (Ser2481)) Polyclonal Antibody, Unconjugated (TMAB-01459) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
9. Sample:
Jurkat (Human) Cell Lysate at 30 μg
MCF-7 (Human) Cell Lysate at 30 μg
Primary: Anti-Phospho-mTOR (Ser2481) (TMAB-01459) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 289 kDa
Observed band size: 289 kDa

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:100; IF: 1:100-500; FCM: 1μg /test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human mTOR around the phosphorylation site of Ser2481

Human

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