Antibody Type: Rabbit Polyclonal

Application: IHC-P,IHC-Fr,ICC/IF,IF,FCM

Reactivity: Human,Rat (predicted:Mouse,Chicken,Pig,Sheep)

IgG

Human,Rat (predicted:Mouse,Chicken,Pig,Sheep)

1. MCF7 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 (Estrogen Receptor alpha) polyclonal Antibody, Unconjugated (TMAB-00631) 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.
2. Blank control (blue line): MCF7 (blue).
Primary Antibody (green line): Rabbit Anti-Estrogen Receptor alpha antibody (TMAB-00631)
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.
Protocol
The cells were fixed with 80% ethanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 30 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature.
3. Paraformaldehyde-fixed, paraffin embedded (Rat ovary); 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 (Estrogen Receptor alpha) Polyclonal Antibody, Unconjugated (TMAB-00631) at 1:400 overnight at 4°C, followed by a conjugated secondary antibody for 20 min and DAB staining.
4. Paraformaldehyde-fixed, paraffin embedded (rat uterus); 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 (Estrogen Receptor alpha) Polyclonal Antibody, Unconjugated (TMAB-00631) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

IHC-P,IHC-Fr,ICC/IF,IF,FCM

IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:50-1:200; IF: 1:100-500; FCM: 1μg /test

Polyclonal

KLH conjugated synthetic peptide: human Estradiol receptor Alpha

Human

2099

Estrogen,ChIP antibodies,Estrogen,Gonadotrophic axis,Estrogen