Anti-Phospho-SIRT1 (Ser47) Polyclonal Antibody is a Rabbit antibody targeting Phospho-SIRT1 (Ser47). Anti-Phospho-SIRT1 (Ser47) Polyclonal Antibody can be used in IHC-P,IHC-Fr,IF,FCM.

IgG

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

1. 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-SIRT1 (Ser47) Polyclonal Antibody, Unconjugated (TMAB-01493) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Tissue/cell: human lung cancer; 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-SIRT1 (Ser47) Polyclonal Antibody, Unconjugated (TMAB-01493) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
3. Blank control (blue line): MCF 7 (fixed with 70% methanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 30 min on ice).
Primary Antibody (green line): Rabbit Anti-Phospho-SIRT1 (Ser47) antibody (TMAB-01493), Dilution: 3 μg/10^5 cells.
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1 μg/test.
4. Paraformaldehyde-fixed, paraffin embedded (rat adrenal gland); 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-SIRT1 (Ser47)) Polyclonal Antibody, Unconjugated (TMAB-01493) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
5. Blank control: HL-60. Primary Antibody (green line): Rabbit Anti-Phospho-SIRT1 (Ser47) antibody (TMAB-01493)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF488
Dilution: 1 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. 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.

IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1ug/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human SirT1 around the phosphorylation site of Ser47

Human

p53 Pathway,Acetylation,Class III / Sir2 class,Obesity,Host Virus Interaction,Other Nuclear Bodies