MA5-14501

antibody from Invitrogen Antibodies

Targeting: ESR1 ER-alpha, Era, ESR, NR3A1

Provider product page for MA5-14501

Western blot

Immunocytochemistry

Immunohistochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [298]
Comments [0]
Validations
Western blot [4]
Immunocytochemistry [3]
Immunohistochemistry [1]
Other assay [6]

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Product number: MA5-14501 - Provider product page
Provider: Invitrogen Antibodies
Product name: Estrogen Receptor alpha Monoclonal Antibody (SP1)
Antibody type: Monoclonal
Antigen: Synthetic peptide
Description: This antibody strongly stains the nucleus of epithelial cells in breast carcinomas. It reacts with ER alpha, 67 kDa. No special pretreatment is required for IHC staining.
Reactivity: Human
Host: Rabbit
Isotype: IgG
Antibody clone number: SP1
Vial size: 500 µL
Concentration: 0.015 mg/mL
Storage: -20° C, Avoid Freeze/Thaw Cycles

ESR1 protein structure - MA5-14501 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Estrogen Receptor (ER) was performed by loading the indicated amounts of recombinant full-length human ER alpha (Product # A15674) and ER beta (Product # A15664) proteins, and 10 µL of PageRuler Prestained Protein Ladder (Product # 26616) per well onto a Novex® 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a PVDF membrane (Product # 88518) using the G2 Blotter (Product # 62288), and blocked with 5% milk in TBST for at least 1 hour at room temperature. ER alpha was detected at ~66 kD (left panel) using an ER rabbit monoclonal antibody (Product # MA5-14501) at a dilution of 1:500 in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-rabbit IgG secondary antibody (Product # 31460) at a dilution of 1:40,000. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080). The blot was then stripped with Restore Plus Western blot stripping buffer (Product # 46430) for at least 15 minutes at room temperature, re-blocked with 5% milk in TBST for at least 1 hour at room temperature, and re-probed with an ER beta polyclonal antibody (Product # PA1-313, right panel) at a dilution of 1:2000 overnight at 4C, followed by an HRP-conjugated goat anti-rabbit IgG secondary antibody (Product # 31460) at a dilution of 1:40,000. Chemiluminescent detection was again performed using SuperSignal West Pico (Product # 34080).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Estrogen Receptor (ER) was performed by loading 20 µg of whole cell lysates (WCL) or nuclear extracts (NE) from the indicated breast cancer cell lines, and 10 µL of PageRuler Prestained Protein Ladder (Product # 26616) per well onto a Novex® 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a PVDF membrane (Product # 88518) using the G2 Blotter (Product # 62288), and blocked with 5% milk in TBST for at least 1 hour at room temp. ER was detected at ~66 kD (top panel) using an ER rabbit monoclonal antibody (Product # MA5-14501) at a dilution of 1:500 in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-rabbit IgG secondary antibody (Product # 31460) at a dilution of 1:40,000. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075). To check for equivalent protein loading in each lane, the blot was stripped with Restore Plus Western blot stripping buffer (Product # 46430) for at least 15 min at room temp, re-blocked with 5% milk in TBST for at least 1 hour at room temp, and re-probed with an HDAC1 polyclonal antibody (Product # PA1-860, bottom panel) at a dilution of 1:1000 overnight at 4C, followed by an HRP-conjugated goat anti-rabbit IgG secondary antibody (Product # 31460) at a dilution of 1:40,000 and chemiluminescent detection. The molecular weight of the endogenous protein detected is consistent with ER alpha.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of Estrogen Receptor using Estrogen Receptor Monoclonal Antibody (Product # MA5-14501) on MCF-7 Cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Estrogen Receptor alpha Monoclonal Antibody (Product # MA5-14501) and a 66kDa band corresponding to Estrogen Receptor alpha was observed across all the cell lines tested with uncharacterized band (*). Whole cell extracts (30 µg lysate) of T-47D (Lane 1), MCF-7 (Lane 2), SK-BR-3 (Lane 3), HepG2 (Lane 4) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Estrogen Receptor (ER, green) in T47D and Hs 578T breast cancer cells. The cells were fixed with 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 15 minutes at room temperature, and blocked with 0.3% BSA in PBS for at least 30 minutes at room temperature. Cells were stained with an ER rabbit monoclonal antibody (Product # MA5-14501) at a dilution of 1:50 in blocking buffer overnight at 4C, and then incubated with a DyLight 488-conjugated goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:500 for at least 1 hour at room temperature. Nuclei (blue) were stained with DAPI (Product # 62247). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Estrogen Receptor (ER, green) in T47D and Hs 578T breast cancer cells. The cells were fixed with 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 15 minutes at room temperature, and blocked with 0.3% BSA in PBS for at least 30 minutes at room temperature. Cells were stained with an ER rabbit monoclonal antibody (Product # MA5-14501) at a dilution of 1:50 in blocking buffer overnight at 4C, and then incubated with a DyLight 488-conjugated goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:500 for at least 1 hour at room temperature. Nuclei (blue) were stained with DAPI (Product # 62247). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Estrogen Receptor alpha was performed using 70% confluent log phase T-47D cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with beta Estrogen Receptor alpha Monoclonal Antibody (Product # MA5-14501) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents merged image for SK-BR-3 cells showing no staining for Estrogen Receptor alpha. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Immunohistochemical staining with ER (upper panel) and H&E staining (lower panel) . Stainings were performed on formalin-fixed, paraffin-embedded tissue sections of invasive ductal breast carcinoma, and photographed before and after preoperative chemotherapy treatment of EFC regimen, by using a Zeiss Axioskop40 Microscope and a Nikon E4500 camera. In ER+ tumor sample (left), tumor cells arranged in glands and clusters still showed active growth after the EFC chemotherapy treatment. Both core needle biopsy specimen and surgical specimen showed diffuse nuclear staining for estrogen receptor (diaminobenzidine chromogen). However, in ER- tumor sample (right), only a small nest of tumor cells remained survival after EFC treatment, with prominent stromal fibrosis and hyalinization. ER staining was negative in both biopsy and surgical specimens. Data are representative of at least three separate experiments. Magnification :x 100 .

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1. Representative immunohistochemical staining of ERalpha and ERbeta in lung adenocarcinoma tissue. All images were captured at magnification, x200. (A) positive nuclear ERalpha staining, (B) positive nuclear ERbeta staining, (C) positive cytoplasmic ERbeta staining, and (D) negative control. ER estrogen receptor.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 The expression of estradiol receptors and progesterone receptors in the uterus of rats after rhFSH treatment. For the detection of ( A ) estradiol receptor and ( B ) progesterone receptor by immunohistochemical staining. The degrees of staining were categorized as follows: (0), no staining; (1), faint staining; (2), moderate staining; and (3), strong staining. Arrows pinpointed the staining of ER or PR. Scale bar = 500 mum. G1: abdominal s.c. injection of normal saline twice daily for 4 days (days 1 to 4); G2: abdominal s.c. injection of rhFSH (1 IU) twice daily for 4 days (days 1 to 4); G3: vaginal s.c. injection of rhFSH (1 IU) twice daily for 4 days (days 1 to 4); G4: abdominal s.c. injection of rhFSH (4 IU) every other day (two injections) (days 1 & 3); G5: vaginal s.c. injection of rhFSH (4 IU) every other day (two injections) (days 1 & 3). Abbreviations: LE: luminal endometrium; GE: glandular endometrium; Myomet: myometrium.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 TET protein expression in response to different steroid hormone treatments in AN3 cells. A representative blot image for the particular weight band is shown next to the graph. The y-axis shows the fold change of protein levels following different treatments compared to control and x-axis shows the different treatment groups. E24, 24h Estrogen; EP24, EP48 and EP72 = both Estrogen + Progesterone for 24, 48 and 72h. Data are presented as mean +- SEM, *p < 0.05 (P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Steroid hormone protein expression in response to different steroid hormone treatments in AN3 cells. A representative blot image for the particular weight band is shown next to the graph. The y-axis shows the fold change of protein levels following different treatments compared to control and x-axis shows the different treatment groups. E24 = 24h Estrogen; EP24, EP48 and EP72 = both Estrogen + Progesterone for 24, 48 and 72h. Data are presented as mean +- SEM. P