ER alpha /NR3A1 in Human Endometrial Cancer Tissue.
ER alpha /NR3A1 was detected in immersion fixed paraffin-embedded sections of human endometrial cancer tissue using Sheep Anti-Human ER alpha /NR3A1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5715) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human ER alpha /NR3A1 by Western Blot.
Western blot shows lysates of MCF‑7 human breast cancer cell line and MDA‑MB‑468 human breast cancer cell line. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human ER alpha /NR3A1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5715) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for ER alpha /NR3A1 at approximately 65 to 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human ER alpha /NR3A1 by Simple WesternTM.
Simple Western lane view shows lysates of MCF‑7 human breast cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for ER alpha /NR3A1 at approximately 66 kDa (as indicated) using 5 µg/mL of Sheep Anti-Human ER alpha /NR3A1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5715) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: ER alpha/NR3A1
ER alpha (Estrogen receptor alpha; also Estradiol receptor and NR3A1) is a 65‑70 kDa member of the NR3 subfamily, nuclear hormone receptor family of proteins. It is widely expressed, and serves as a strong activator of estrogen-responsive genes. ER alpha is normally quiescent and bound to heat-shock proteins and immunophilins. Following beta -estradiol binding, it becomes activated, either homodimerizes or heterodimerizes with ER beta, and binds to DNA with multiple coactivators. Human ER alpha is 595 amino acids (aa) in length, and contains a DNA binding region (aa 185‑250), three NLSs (aa 256‑260; 266‑271; 299‑303), a steroid‑binding site (aa 351‑543), a dimerization motif (aa 497‑518), and an O-GlcNAc attachment around Thr575. Major phosphorylation sites exist at Tyr537, Ser167 and Ser118.
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