MAM was purchased from Midwest Research Institute (Kansas City, MO, USA). Chloral hydrate was sourced from Sigma-Aldrich (St. Louis, MO, USA), while heparin was from PSS World Medical, Inc (Jacksonville, FL, USA). Mifepristone (RU-486) and fulvestrant (ICI 182,780) were purchased from Tocris (Minneapolis, MN, USA). The Rat Estradiol ELISA kit was obtained from Calbiotech (Spring Valley, CA, USA) and the Progesterone ELISA kit was purchased from Abnova (Walnut, CA, USA). All other chemicals and reagents were of either analytical or laboratory grade, and purchased from standard suppliers.
Fulvestrant ici182780
Manufactured by Bio-Techne
Sourced in United States
Fulvestrant (ICI182780) is a synthetic estrogen receptor antagonist. It binds to the estrogen receptor and blocks its activation, thereby inhibiting the effects of estrogen in target tissues.
Automatically generated - may contain errors
Lab products found in correlation
4 hydroxytamoxifen 4 oht, by Merck Group (2 mentions)
Chloral hydrate, by Merck Group (1 mentions)
Total p27, by Cell Signaling Technology (1 mentions)
Dasatinib, by Selleck Chemicals (1 mentions)
Cyclin d1, by Cell Signaling Technology (1 mentions)
Total egfr, by Cell Signaling Technology (1 mentions)
Phospho src tyr 527, by Cell Signaling Technology (1 mentions)
Total erbb2, by Merck Group (1 mentions)
Phospho src tyr416, by Cell Signaling Technology (1 mentions)
Total akt, by Cell Signaling Technology (1 mentions)
Phospho akt ser473, by Cell Signaling Technology (1 mentions)
Phospho egfr tyr1068, by Cell Signaling Technology (1 mentions)
17β estradiol e2, by Merck Group (1 mentions)
Everolimus rad001, by Bio-Techne (1 mentions)
17β estradiol e, by Merck Group (1 mentions)
4 protocols using fulvestrant ici182780
1
Hormone Quantification Assay Protocol
2
Immunoblotting Analysis of Signaling Proteins
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Primary antibodies against phospho-srctyr416, phospho-srctyr527, src, phospho-ERK1/2 (p42/44), phospho-p90rsk, phospho-AKTser473, total-AKT, cyclin D1, phospho-EGFRtyr1068, total-EGFR and total p27 were purchased from Cell Signalling; total-ER and phospho-p27ser10 from Santa Cruz; total ERK1/2 and Actin (Sigma Aldrich) and PgR (Novacastra). Total-ERBB2 and phospho-ERBB2Tyr1248 were purchased from Millipore. Secondary horseradish peroxidase antibodies were obtained from Dako. 17-β-estradiol (E2) and 4-hydroxytamoxifen (4-OHT) were purchased from Sigma-Aldrich, UK; fulvestrant (ICI182780) from Tocris Bioscience and dasatinib from Selleckchem.
3
Fulvestrant and Everolimus Treatment Protocol
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Fulvestrant/ICI 182,780 and Everolimus/RAD001 were purchased from Tocris, Minneapolis, MN, USA. Fulvestrant and Everolimus were dissolved in 100% ethanol to a stock concentration of 10 mM and stored at −20 °C. Working stocks were prepared fresh at the time of treatment. Antibodies used are listed in Table S1 .
4
Immunoblotting and ChIP Assay Protocol
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Following antibodies were used for immunoblotting: pESR1ser167 (CST cat-5587, 1:1000), pESR1ser118 (CST cat-2511, 1:1000), total-ESR1 (Santa Cruz sc8002, 1:800 or Novacastra (NCL-ER-6F11), 1:1000), total-FOXA1 (Abcam Ab23738, 1:1000) total-PGR (Novocastra NCL-L-PGR, 1:500 or Santa Cruz sc-538, 1:200), pERBB2 (CST-2243, 1:1000), total-ERBB2 (CST-4290, 1:1000), pEGFR (CST-3777, 1:1000), total-EGFR (CST-2232, 1:1000), pAKTser437 (CST-9271, 1:1000), total-AKT (CST-9272, 1:1000), pERK1/2 (Sigma-Aldrich, 1:2000), total-ERK1/2 (CST-9102, 1:1000), TFF1 (Santa Cruz sc28925, 1:200), RARA (Abcam Ab39971, 1:1000), cathepsin D (CTSD) (Abcam Ab6313, 1:2000), actin (Abcam Ab6276, 1:10,000), and tubulin (Sigma T-9026, 1:2000). Secondary antibodies (horseradish peroxidase-linked, 1:2000) were obtained from Dako. For ChIP, the following antibodies were used: ESR1 (Santa Cruz sc543), CBP (Santa Cruz sc369), and FOXA1 (Abcam Ab23738). 17-β-estradiol (E) and 4-hydroxytamoxifen (4-OHT) were purchased from Sigma-Aldrich and fulvestrant (ICI182780) from Tocris Bioscience.
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