Estrogen Receptor alpha + beta Rabbit Polyclonal Antibody

• Catalog Number: orb10615
• Category: Antibodies
• Description: Estrogen Receptor alpha + beta Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Mouse, Porcine
• Reactivity: Human, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human Estradiol Receptor alpha + beta (201-300/595aa)
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100, IF=1:100-500, Flow-Cyt=1ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 66 kDa
• Target: ESR1
• UniProt ID: P03372
• RRID: AB_10750914
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: Estrogen Receptor alpha/beta; Atherosclerosis, sus
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control (blue line): MCF7 (blue). Primary Antibody (green line): Rabbit Anti-Estrogen Receptor alpha + beta antibody (orb10615), Dilution: 5 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. Acquisition of 20000 events was performed.

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 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (Estrogen Receptor alpha + beta) Polyclonal Antibody, Unconjugated (orb10615) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Protein: Brain (Rat) lysate at 30 ug, Colon (Rat) lysates, 30 ug, Primary: Anti-ER-alpha/beta (orb10615) at 1:200, Secondary: HRP conjugated Goat-Anti-Rabbit IgG at 1:3000, Predicted band size: 58kD, 66kD, Observed band size: 58kD, ER-alpha: 66kD, ER-beta: 58kD.

Tissue/Cell: human endometrium tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-ER-alpha/beta Polyclonal Antibody, Unconjugated (orb10615) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

Tissue/Cell: HUVEC 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 + beta) Polyclonal Antibody, Unconjugated (orb10615) 1:100, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (orb868805) at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.