Anti-Phospho-AR/Androgen receptor (Ser578) Polyclonal Antibody is a Rabbit antibody targeting Phospho-AR/Androgen receptor (Ser578). Anti-Phospho-AR/Androgen receptor (Ser578) Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

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

1. Blank control (Black line): U87MG (Black).
Primary Antibody (green line): Rabbit Anti-phospho-Androgen Receptor (Ser578) antibody (TMAB-10332)
Dilution: 1 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF647
Dilution: 1 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 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. Acquisition of 10,000 events was performed.
2. Tissue/cell: human prostate tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-phospho-Androgen Receptor(Ser580) Polyclonal Antibody, Unconjugated (TMAB-10332) 1: 200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
3. Tissue/cell: human prostate tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-AIS Polyclonal Antibody, Unconjugated (TMAB-10332) 1:500, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
4. Sample:
Muscle (Mouse) Lysate at 40 μg
Uterus (Mouse) Lysate at 40 μg
Ovary (Rat) Lysate at 40 μg
DU145 (Human) Cell Lysate at 40 μg
Primary: Anti-phospho-Androgen Receptor (Ser578) (TMAB-10332) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 101 kD
Observed band size: 101 kD
5. Paraformaldehyde-fixed, paraffin embedded (mouse prostate); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30 min; Incubation with (phospho-AR/Androgen receptor (Ser578)) Polyclonal Antibody, Unconjugated (TMAB-10332) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 3μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human Androgen Receptor around the phosphorylation site of Ser578

Human

AR

Androgen receptor

Steroid hormone receptors are ligand-activated transcription factors that regulate eukaryotic gene expression and affect cellular proliferation and differentiation in target tissues. Transcription factor activity is modulated by bound coactivator and corepressor proteins. Transcription activation is down-regulated by NR0B2. Activated, but not phosphorylated, by HIPK3.