Sample: Lung (Mouse) Lysate at 40 ug
Primary: Anti-phospho-AKT3 (Ser472) (SL5209R) at 1/300 dilution
Secondary: HRP conjugated Goat-Anti-rabbit IgG (SL0295G-HRP) at 1/5000 dilution
Predicted band size: 56 kD
Observed band size: 56 kD
Sample:
Lane 1: Huvec (Human) Cell Lysate at 30 ug
Lane 2: A431 (Human) Cell Lysate at 30 ug
Primary: Anti-phospho-AKT3 (Ser472) (SL5209R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 60 kD
Observed band size: 58 kD
Sample:
brain (Mouse) Lysate at 40 ug
Primary: Anti-phospho-AKT3 (Ser472) (SL5209R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 56 kD
Observed band size: 56 kD
Sample:
Lung (Mouse) Lysate at 40 ug
Primary: Anti-phospho-AKT3 (Ser472) (SL5209R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 56 kD
Observed band size: 56 kD
Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-AKT3 (Ser472)) Polyclonal Antibody, Unconjugated (SL5209R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-AKT3 (Ser472)) Polyclonal Antibody, Unconjugated (SL5209R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-AKT3 (Ser472)) Polyclonal Antibody, Unconjugated (SL5209R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-AKT3 (Ser472)) Polyclonal Antibody, Unconjugated (SL5209R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Tissue/cell: human lung carcinoma;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,SLC0005) at 37鈩� for 20 min;
Incubation: Anti-phospho-AKT3(Ser472) Polyclonal Antibody, Unconjugated(SL5209R) 1:200, overnight at 4掳C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated (SL0295G-Cy3)used at 1:200 dilution for 40 minutes at 37掳C. DAPI(5ug/ml,blue,SLC0033) was used to stain the cell nuclei
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