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Erα f 10

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ERα (F-10) is a mouse monoclonal antibody that recognizes the estrogen receptor alpha (ERα) protein. It can be used to detect ERα expression in various experimental applications.

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Lab products found in correlation

Alpha tubulin ab80779, by Abcam (2 mentions) Psmc2 mss1 104, by Enzo Life Sciences (2 mentions) Psmd1 c 7, by Santa Cruz Biotechnology (2 mentions) Np0009, by Thermo Fisher Scientific (2 mentions) Beta actin n 21, by Santa Cruz Biotechnology (2 mentions) Odyssey clx imaging machine, by LI COR (2 mentions) Image studio software, by LI COR (2 mentions) Image lab software, by Bio-Rad (2 mentions) Protease inhibitor cocktail, by Roche (2 mentions) Chemidoc mp system, by Bio-Rad (2 mentions) Estetrol, by Merck Group (1 mentions) Parp1 2 sc 7150, by Santa Cruz Biotechnology (1 mentions) 17β estradiol, by Merck Group (1 mentions) β actin 13e5 rabbit mab, by Cell Signaling Technology (1 mentions) Supersignal west pico plus chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions) Trans blot turbo rta mini nitrocellulose transfer kit, by Bio-Rad (1 mentions) Parp 46d11 rabbit mab, by Cell Signaling Technology (1 mentions) Rb if8, by Santa Cruz Biotechnology (1 mentions) Plk1 208g4, by Cell Signaling Technology (1 mentions) Ripa lysis buffer, by Santa Cruz Biotechnology (1 mentions)

5 protocols using erα f 10

1

Quantitative Immunoblotting of PSMC2, PSMD2, and ERα

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For PSMC2 and PSMD2 immunoblotting, cells were lysed in HENG buffer (50mM Hepes-KOH pH 7.9, 150mM NaCl, 2mM EDTA pH 8.0, 20mM sodium molybdate, 0.5% Triton X-100, 5% glycerol), with protease inhibitor cocktail (Roche Diagnostics #11836153001). Protein concentration was determined by the BCA assay (Thermo-Fisher Scientific #23227), and proteins were resolved on SDS-PAGE for immunoblot analysis. Antibodies against the following human proteins were used: alpha-Tubulin (ab80779; Abcam), PSMC2 (MSS1–104; Enzo Life Sciences) and PSMD1 (C-7; Santa-Cruz). Visualization was performed using the ChemiDoc MP System (Bio-Rad), and ImageLab Software (Bio-Rad) was used to quantify relative band intensities. For ERα immunonblotting, cells were lysed with a mix of 4X protein loading buffer (Li-Cor 928–40004) and 10X NuPAGE sample reducing agent (Life Technologies NP0009). Protein concentration was normalized by cell counting, and proteins were resolved on SDS-PAGE. Antibodies against the following human proteins were used: beta-Actin (N-21; Santa Cruz), ERα (F-10; Santa Cruz). Visualization was performed using the Odyssey CLx imaging machine (Li-Cor), and Image Studio Software (Li-Cor) was used to quantify the relative intensities.
Ben-David U., Siranosian B., Ha G., Tang H., Oren Y., Hinohara K., Strathdee C.A., Dempster J., Lyons N.J., Burns R., Nag A., Kugener G., Cimini B., Tsvetkov P., Maruvka Y.E., O’Rourke R., Garrity A., Tubelli A.A., Bandopadhayay P., Tsherniak A., Vazquez F., Wong B., Birger C., Ghandi M., Thorner A.R., Bittker J.A., Meyerson M., Getz G., Beroukhim R, & Golub T.R. (2018). Genetic and transcriptional evolution alters cancer cell line drug response. Nature, 560(7718), 325-330.
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2

Quantitative Immunoblotting of PSMC2, PSMD2, and ERα

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For PSMC2 and PSMD2 immunoblotting, cells were lysed in HENG buffer (50mM Hepes-KOH pH 7.9, 150mM NaCl, 2mM EDTA pH 8.0, 20mM sodium molybdate, 0.5% Triton X-100, 5% glycerol), with protease inhibitor cocktail (Roche Diagnostics #11836153001). Protein concentration was determined by the BCA assay (Thermo-Fisher Scientific #23227), and proteins were resolved on SDS-PAGE for immunoblot analysis. Antibodies against the following human proteins were used: alpha-Tubulin (ab80779; Abcam), PSMC2 (MSS1–104; Enzo Life Sciences) and PSMD1 (C-7; Santa-Cruz). Visualization was performed using the ChemiDoc MP System (Bio-Rad), and ImageLab Software (Bio-Rad) was used to quantify relative band intensities. For ERα immunonblotting, cells were lysed with a mix of 4X protein loading buffer (Li-Cor 928–40004) and 10X NuPAGE sample reducing agent (Life Technologies NP0009). Protein concentration was normalized by cell counting, and proteins were resolved on SDS-PAGE. Antibodies against the following human proteins were used: beta-Actin (N-21; Santa Cruz), ERα (F-10; Santa Cruz). Visualization was performed using the Odyssey CLx imaging machine (Li-Cor), and Image Studio Software (Li-Cor) was used to quantify the relative intensities.
Ben-David U., Siranosian B., Ha G., Tang H., Oren Y., Hinohara K., Strathdee C.A., Dempster J., Lyons N.J., Burns R., Nag A., Kugener G., Cimini B., Tsvetkov P., Maruvka Y.E., O’Rourke R., Garrity A., Tubelli A.A., Bandopadhayay P., Tsherniak A., Vazquez F., Wong B., Birger C., Ghandi M., Thorner A.R., Bittker J.A., Meyerson M., Getz G., Beroukhim R, & Golub T.R. (2018). Genetic and transcriptional evolution alters cancer cell line drug response. Nature, 560(7718), 325-330.
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3

Estradiol, Estetrol, and DMSO Examination

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First, 17β-estradiol, estetrol, and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Ethanol was acquired from Winkler (Santiago, Chile). The primary antibodies were PARP 1/2 (sc-7150) and ERα (F-10) (Santa Cruz, CA, USA). Progesterone receptor (6A1) (Cell Signaling, MA, USA), ERβ (Abx121395), and β-actin (Abx133823) (Abbexa, Cambridge, UK).
Patiño-García D., Palomino J., Pomés C., Celle C., Torres-Estay V, & Orellana R. (2023). Estetrol Increases Progesterone Genetic Response without Triggering Common Estrogenic Effects in Endometriotic Cell Lines and Primary Cultures. Biomedicines, 11(4), 1169.
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4

Western Blot Analysis of Cell Signaling

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Cells were lysed with RIPA lysis buffer (sc-24948, Santa Cruz Biotechnology, Inc., Dallas, TX) according to the protocol supplied. Whole cell lysates (30 μg) were separated by SDS-PAGE, transferred to nitrocellulose through Trans-Blot Turbo RTA Mini Nitrocellulose Transfer Kit (Bio-Rad, Hercules, CA, USA). Membranes were subjected to immunoblot analyses using primary antibodies against phosphorylated RB (Ser780) #8180 1:1,000 (Cell Signaling Technology, Danvers, MA), phosphorylated RB (Ser807/811) #8516 1:1,000 (Cell Signaling Technology), RB (IF-8) #sc102 1:1,000 (Santa Cruz Biotechnology, Inc.), Erα (F-10) #sc-8002 1:200 (Santa Cruz Biotechnology, Inc.), PLK1 (208G4) #4513 (Cell Signaling Technology) 1:1,000, β-actin (13E5) rabbit mAb #4970 (Cell Signaling Technology) 1:5,000, PARP (46d11) rabbit mAb #9532 (Cell Signaling Technology), GAPDH #sc-32233 1:10,000 (Santa Cruz Biotechnology, Inc.), HRP-conjugated anti-rabbit and anti-mouse were used as secondary antibodies (Bio-Rad). Immunoreactive proteins were visualized by enhanced chemiluminescence using SuperSignal West Pico PLUS Chemiluminescent Substrate (Thermo Fisher Scientific, Waltham, MA). Membranes were cut horizontally to probe with multiple antibodies. Films were imaged using Brother MFCL2710DW (Brother) at 300 dpi.
Guerrero-Zotano Á., Belli S., Zielinski C., Gil-Gil M., Fernandez-Serra A., Ruiz-Borrego M., Ciruelos Gil E.M., Pascual J., Muñoz-Mateu M., Bermejo B., Margeli Vila M., Antón A., Murillo L., Nissenbaum B., Liu Y., Herranz J., Fernández-García D., Caballero R., López-Guerrero J.A., Bianco R., Formisano L., Turner N, & Martín M. (2023). CCNE1 and PLK1 Mediate Resistance to Palbociclib in HR+/HER2− Metastatic Breast Cancer. Clinical Cancer Research, 29(8), 1557-1568.
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5

Optimized Antibody Detection Protocol

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All chemicals were purchased from Sigma-Aldrich (Taufkirchen, Germany) unless stated otherwise. Furthermore, Dulbecco's phosphate-buffered saline (DPBS) (PAN-Biotech, Aidenbach, Germany) and Bisphenol A/C (BPA/C) (Angene, London, UK) was used. The following antibodies were used: β-actin (AC-15), α-tubulin, cytochrome C (EPR1327), mitofilin (2E4AD5) (all from Abcam, Cambridge, UK) and ERα (F-10) (Santa Cruz Biotechnology, Heidelberg, Germany).
Padberg F., Tarnow P., Luch A, & Zellmer S. (2019). Minor structural modifications of bisphenol A strongly affect physiological responses of HepG2 cells. Archives of toxicology, 93(6).
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