bs-0174R [Primary Antibody]
Rabbit  Anti-Estrogen Receptor alpha + beta Polyclonal Antibody
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Host: Rabbit

Target Protein: Estrogen Receptor alpha + beta

IR: Immunogen Range:201-300/595

Clonality: Polyclonal

Isotype: IgG

Entrez Gene: 2099

Swiss Prot: P03372

Source: KLH conjugated synthetic peptide derived from human Estradiol Receptor alpha + beta:201-300/595 

Purification: affinity purified by Protein A

Storage: 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Background: The estrogen receptor (ER) is a 66 kDa protein that mediates the actions of estrogens in estrogen responsive tissues. It is a member of a large superfamily of nuclear hormone receptors that function as ligand activated transcription factors. The ER gene consists of more than 140 kb of genomic DNA divided into 8 exons. These translate into a protein with six functionally discrete domains, labeled A through F. A second form of the estrogen receptor, ER beta has recently been described. The ER is an important regulator of growth and differentiation in the mammary gland. Presence of ER in breast tumors indicates an increased likelihood of response to anti estrogen (e.g. tamoxifen) therapy.

Size: 100ul

Concentration: 1mg/ml

Applications: WB=1:500-2000,IHC-P=1:100-500,IHC-F=1:100-500,Flow-Cyt=1ug/Test,ICC=1:100,IF=1:100-500,ELISA=1:5000-10000

Cross Reactive Species: Human,Rat (predicted: Mouse,Pig)

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES
Protein: Brain(Rat) lysate at 30ug; Colon(Rat) lysates, 30ug; Primary: Anti-ER-alpha/beta (bs-0174R) at 1:200 Secondary: HRP conjugated Goat-Anti-Rabbit IgG(bse-0295G) at 1: 3000 Predicted band size : 58kD,66kD Observed band size : 58kD ER-alpha: 66kD; ER-beta: 58kD;
Paraformaldehyde-fixed, paraffin embedded (rat uterus); 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 (Estrogen Receptor alpha + beta) Polyclonal Antibody, Unconjugated (bs-0174R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Tissue/cell: human endometrium tissue; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min; Incubation: Anti-ER-alpha/beta Polyclonal Antibody, Unconjugated(bs-0174R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
Tissue/cell: HUVEC cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (Estrogen Receptor alpha + beta ) Polyclonal Antibody, Unconjugated (bs-0174R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-FITC) at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
Blank control (blue line): MCF7 (blue). Primary Antibody (green line): Rabbit Anti-Estrogen Receptor alpha + beta antibody (bs-0174R) Dilution: 5μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC Dilution: 1μg /test. Protocol The cells were fixed with 80% ethanol (Overnight at 4℃) 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. Acquisition of 20,000 events was performed.
 
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