Estrogen receptor alpha Rabbit Polyclonal Antibody

• Catalog Number: orb10614
• Category: Antibodies
• Description: Estrogen receptor alpha Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, WB
• Predicted Reactivity: Mouse
• Reactivity: Human, Mouse
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human ER-Alpha (501-595/595aa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, ICC/IF=1:100, Flow-Cyt=1μg/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 66 kDa
• Target: ESR1
• UniProt ID: P03372
• RRID: AB_10924663
• 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: Estradiol receptor; Estradiol Receptor-alpha; Estr
• 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 antibody (orb10614), 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. Acquisition of 20000 events was performed.

Sample: K562 (Human) cell Lysate at 40 ug, Primary: Anti-Estrogen receptor alpha (orb10614) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-RabbitIgG at 1/20000 dilution, Predicted band size: 66kD, Observed band size: 63kD.

Sample: Lymph node (Mouse) Lysate at 40 ug, Primary: Anti-Estrogen receptor alpha (orb10614) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 66 kD, Observed band size: 66 kD.

Tissue/Cell: 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 (orb10614) 1:100, 90 minutes at 37°C, followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.