Anti-Phospho-AKT3 (Ser472) Polyclonal Antibody is a Rabbit antibody targeting Phospho-AKT3 (Ser472). Anti-Phospho-AKT3 (Ser472) Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF.

IgG

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

1. Paraformaldehyde-fixed, paraffin embedded (Mouse 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-AKT3 (Ser472)) Polyclonal Antibody, Unconjugated (TMAB-01384) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
2. Tissue/cell: human lung carcinoma;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-phospho-AKT3 (Ser472) Polyclonal Antibody, Unconjugated (TMAB-01384) 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated used at 1:200 dilution for 40 minutes at 37°C. DAPI (5 μg/ml,blue) was used to stain the cell nucleus.
3. 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-AKT3 (Ser472)) Polyclonal Antibody, Unconjugated (TMAB-01384) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
4. Sample:
Lane 1: Huvec (Human) Cell Lysate at 30 μg
Lane 2: A431 (Human) Cell Lysate at 30 μg
Primary: Anti-phospho-AKT3 (Ser472) (TMAB-01384) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 60 kDa
Observed band size: 58 kDa
5. Paraformaldehyde-fixed, paraffin embedded (mouse 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-AKT3 (Ser472)) Polyclonal Antibody, Unconjugated (TMAB-01384) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
6. 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-AKT3 (Ser472)) Polyclonal Antibody, Unconjugated (TMAB-01384) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500

Polyclonal

KLH conjugated Synthesised phosphopeptide: human AKT3 around the phosphorylation site of Ser472

Human