ERa/NR3A1 was detected in immersion fixed paraffin-embedded sections of human endometrial cancer tissue using Sheep Anti-Human ERa/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 ERa/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 ERa/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 Western^TM.

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 ERa/NR3A1 at approximately 66 kDa (as indicated) using 5 µg/mL of Sheep Anti-Human ERa/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.

Applications for Human ER alpha/NR3A1 Antibody

Application

Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human endometrial cancer tissue subjected to Antigen Retrieval Reagent-Basic (Catalog # CTS013)

Simple Western

5 µg/mL
Sample: MCF‑7 human breast cancer cell line

Western Blot

0.5 µg/mL
Sample: MCF-7 human breast cancer cell line and MDA-MB-468 human breast cancer cell line

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Long Name

Estrogen Receptor alpha

Alternate Names

ESR1, NR3A1

Entrez Gene IDs

2099 (Human)

Gene Symbol

ESR1

UniProt

P03372

Additional ER alpha/NR3A1 Products

Product Documents for Human ER alpha/NR3A1 Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Human ER alpha/NR3A1 Antibody

For research use only

Citations
Reviews

Human ER alpha/NR3A1 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF5715

Key Product Details

Species Reactivity

Validated:

Cited:

Applications

Validated:

Cited:

Label

Antibody Source

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Product Specifications

Immunogen

Specificity

Clonality

Host

Isotype

Scientific Data Images for Human ER alpha/NR3A1 Antibody

ER alpha/NR3A1 in Human Endometrial Cancer Tissue.