Anti-Phospho-ER alpha (S118) ESR1 Rabbit Monoclonal Antibody

Western blot analysis of ER/ESR1 (Phospho-S118) using anti-ER/ESR1 (Phospho-S118) antibody (P00057).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: rat brain tissue lysates,
Lane 2: rat C6 whole cell lysates,
Lane 3: mouse brain tissue lysates,
Lane 4: mouse Neuro-2a whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-ER/ESR1 (Phospho-S118) antigen affinity purified monoclonal antibody (P00057) at 1:500 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for ER/ESR1 (Phospho-S118) at approximately 66 kDa. The expected band size for ER/ESR1 (Phospho-S118) is at 66 kDa.

All lanes use the Antibody at 1:1K dilution for 1 hour at room temperature.

Immunofluorescent analysis of MCF7 cells treated with EGF, using Phospho-ER alpha (S118) Antibody.

Western blot analysis of Phospho-ER alpha (S118) expression in (1) MCF7 cell lysate; (2) MCF7 cell lysate treated with b-Estradiol and EGF.

IHC analysis of p-ESR using anti-p-ESR antibody (P00057).
p-ESR was detected in a paraffin-embedded section of rat uterus tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-p-ESR Antibody (P00057) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of p-ESR using anti-p-ESR antibody (P00057).
p-ESR was detected in a paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-p-ESR Antibody (P00057) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of p-ESR using anti-p-ESR antibody (P00057).
p-ESR was detected in a paraffin-embedded section of human prostatic hyperplasia tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-p-ESR Antibody (P00057) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.