17β-estradiol (E2), DMEM (with and without phenol red), and fetal calf serum were purchased from Sigma-Aldrich (St. Louis, MO, USA). Bradford protein assay kit, as well as anti-mouse and anti-rabbit secondary antibodies, were obtained from Bio-Rad (Hercules, CA, USA). Antibodies against ERα (H-C20, rabbit), Bcl-2 (C2, mouse), cathepsin D (H75, rabbit), pS2 (FL-84, rabbit) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA); anti-phospho ERα (Ser118, mouse) antibody was obtained from Cell Signaling; anti-vinculin (mouse) antibody was purchased from Sigma-Aldrich (St. Louis, MO, USA). Chemiluminescence reagent for Western blotting was obtained from BioRad Laboratories (Hercules, CA, USA). Fulvestrant (i.e., ICI182,780) was purchased by Tocris (Bristol, UK), Cycloheximide (CHX) was purchased from Sigma-Aldrich (St. Louis, MO, USA). FDA-approved drug library as well as palbociclib, ribociclib, and abemaciclib were purchased by Selleck Chemicals (Houston, TX, USA). PolarScreen™ ERα Competitor Assay Kit, Green (A15882) was acquired from Thermo Scientific (Waltham, MA, USA). All the other products were from Sigma-Aldrich. Analytical- or reagent-grade products were used without further purification. The identities of all the used cell lines were verified by STR analysis (BMR Genomics, Padova, Italy).
Chemiluminescence reagent for western blotting
Manufactured by Bio-Rad
Sourced in United States
Chemiluminescence reagent for Western blotting is a laboratory product designed to facilitate the detection and quantification of proteins in Western blot analysis. The reagent generates a luminescent signal upon reaction with the enzyme-labeled antibodies used in the Western blotting technique, allowing for the visualization and analysis of target proteins.
Automatically generated - may contain errors
Lab products found in correlation
Bradford protein assay kit, by Bio-Rad (6 mentions)
Fetal calf serum (fcs), by Merck Group (5 mentions)
Anti mouse and anti rabbit secondary antibodies, by Bio-Rad (5 mentions)
Dmem with and without phenol red, by Merck Group (3 mentions)
Anti vinculin antibody, by Merck Group (3 mentions)
Mouse anti vinculin antibody, by Merck Group (2 mentions)
Fulvestrant i e ici182 780, by Bio-Techne (2 mentions)
17β estradiol, by Merck Group (2 mentions)
Faslodex, by Bio-Techne (2 mentions)
Ribociclib, by Selleck Chemicals (1 mentions)
Abemaciclib, by Selleck Chemicals (1 mentions)
Palbociclib, by Selleck Chemicals (1 mentions)
Cycloheximide (chx), by Merck Group (1 mentions)
Bt474, by American Type Culture Collection (1 mentions)
Anti mouse secondary antibodies, by Bio-Rad (1 mentions)
Mda mb 231, by American Type Culture Collection (1 mentions)
Anti akt, by Cell Signaling Technology (1 mentions)
Anti igf 1r, by Cell Signaling Technology (1 mentions)
Anti goat secondary antibody sc 2020, by Santa Cruz Biotechnology (1 mentions)
Anti phospho akt, by Cell Signaling Technology (1 mentions)
6 protocols using chemiluminescence reagent for western blotting
1
Estrogen Receptor Signaling Protocol
2
Evaluation of Estrogen Receptor and Ubiquitin in BT-474 and MDA-MB-231 Cells
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BT-474 and MDAMB231 were purchased by ATCC (Manassas, VA, USA). DMEM (with and without phenol red) and fetal calf serum were purchased from Sigma-Aldrich (St. Louis, MO, USA). Bradford protein assay kit, as well as anti-mouse secondary antibodies, were obtained from Bio-Rad (Hercules, CA, USA). Antibodies against ERα (F-10, mouse) and ubiquitin (P4D1, mouse) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA); anti-vinculin (mouse) antibody was purchased from Sigma-Aldrich (St. Louis, MO, USA). Chemiluminescence reagent for Western blotting was obtained from BioRad Laboratories (Hercules, CA, USA). Fulvestrant (i.e., ICI182,780) was purchased by Tocris (USA). All the other products were from Sigma-Aldrich. Analytical- or reagent-grade products were used without further purification. The identity of the BT-474 and MDAMB231 cell lines was verified by STR analysis (BMR Genomics, Italy).
3
Breast Cancer Cell Line Characterization
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17β-estradiol, DMEM (with and without phenol red) and fetal calf serum were purchased from Sigma-Aldrich (St. Louis, MO). The Bradford protein assay kit, as well as anti-mouse and anti-rabbit secondary antibodies, were obtained from Bio-Rad (Hercules, CA). Antibodies against ERα (HC-20 rabbit; F-10 mouse), cyclin D1 (H-295 rabbit), Bcl-2 (C2 mouse), progesterone receptor (C20 rabbit), cathepsin D (H75 rabbit), pS2 (FL-84 rabbit), dynamin II (C-18 goat), p62/SQSTM (D-3 mouse) and anti-goat secondary antibody (sc-2020) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA) and anti-vinculin, anti-tubulin and anti-LC3 antibodies were purchased from Sigma-Aldrich (St. Louis, MO). Anti-phospho-AKT, anti-AKT, anti-phospho-S118 ERα, and anti-IGF-1R were purchased from Cell Signaling Technology, Inc. (Beverly, MA). Anti-biotin-HRP was purchased from Thermo Fisher Scientific (Walthman, MA, USA). Chemiluminescence reagent for Western blotting was obtained from BioRad Laboratories (Hercules, CA, USA). Dynole 2–24 was purchased from Abcam (USA). All other products were from Sigma-Aldrich. Analytical- or reagent-grade products, without further purification, were used. The identities of all of the cell lines that were used (i.e., human breast carcinoma cells [MCF-7; T47D-1]) were verified by STR analysis (BMR Genomics, Italy).
4
Estrogen Receptor Signaling Pathway Protocol
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17β-Estradiol (E2), Dulbecco’s modified Eagle’s medium (with and without phenol red) and fetal calf serum were purchased from Sigma-Aldrich (St. Louis, MO). A Bradford protein assay kit and anti-mouse and anti-rabbit secondary antibodies were obtained from Bio-Rad (Hercules, CA, USA). Antibodies against ERα (HC-20, rabbit), cyclin D1 (H-295, rabbit), Bcl-2 (C2, mouse), progesterone receptor (C20, rabbit), cathepsin D (H75, rabbit), and pS2 (FL-84, rabbit) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-vinculin antibody was purchased from Sigma-Aldrich (St. Louis, MO, USA). Chemiluminescence reagent for Western blotting was obtained from Bio-Rad Laboratories (Hercules, CA, USA). Fulvestrant (i.e., faslodex or ICI 182,780), AZD9496 and brilanestrant (GDC) were purchased from Tocris (Bristol, UK). Telaprevir (VX-950) was purchased from Selleck Chemicals (Houston, TX, USA). All the other products were obtained from Sigma-Aldrich. Analytical- or reagent-grade products were used without further purification.
5
Breast and Cervical Cancer Cell Culture
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Cell culture and reagents.
17β-estradiol (E2), epidermal growth factor (EGF), 4OH-tamoxifen (Tam), DMEM (with and without phenol red), fetal calf serum, charcoal stripped fetal calf serum (DCC) and the palmitoylacyl-transferase (PAT) inhibitor 2-bromohexadecanoic acid (2-bromo-palmitate; 2-Br) [IC 50 of ~4 µM] (Varner et al., 2003) were purchased from Sigma-Aldrich (St. Louis, MO). Bradford protein assay kit as well as anti-mouse and anti-rabbit secondary antibodies were obtained from Bio-Rad (Hercules, CA). Antibodies against ERα (F-10 mouse -for WB), pS2 (FL-84 rabbit) and cathepsin D (H75 rabbit) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA); anti-vinculin antibody was from Sigma-Aldrich (St. Louis, MO). All other antibodies were purchased by Cell Signalling Technology Inc. (Beverly, MA, USA). Chemiluminescence reagent for Western blotting was obtained from BioRad Laboratories (Hercules, CA, USA). Nano-Glo® Endurazine TM was purchased from Promega (Promega, Madison, MA, USA). All the other products were from Sigma-Aldrich. Analytical-or reagent-grade products were used without further purification. The identities of all the used cell lines [i.e., human breast carcinoma cells (MCF-7 and T47D-1) and human cervix carcinoma cells (HeLa)] were verified by STR analysis (BMR Genomics, Italy).
17β-estradiol (E2), epidermal growth factor (EGF), 4OH-tamoxifen (Tam), DMEM (with and without phenol red), fetal calf serum, charcoal stripped fetal calf serum (DCC) and the palmitoylacyl-transferase (PAT) inhibitor 2-bromohexadecanoic acid (2-bromo-palmitate; 2-Br) [IC 50 of ~4 µM] (Varner et al., 2003) were purchased from Sigma-Aldrich (St. Louis, MO). Bradford protein assay kit as well as anti-mouse and anti-rabbit secondary antibodies were obtained from Bio-Rad (Hercules, CA). Antibodies against ERα (F-10 mouse -for WB), pS2 (FL-84 rabbit) and cathepsin D (H75 rabbit) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA); anti-vinculin antibody was from Sigma-Aldrich (St. Louis, MO). All other antibodies were purchased by Cell Signalling Technology Inc. (Beverly, MA, USA). Chemiluminescence reagent for Western blotting was obtained from BioRad Laboratories (Hercules, CA, USA). Nano-Glo® Endurazine TM was purchased from Promega (Promega, Madison, MA, USA). All the other products were from Sigma-Aldrich. Analytical-or reagent-grade products were used without further purification. The identities of all the used cell lines [i.e., human breast carcinoma cells (MCF-7 and T47D-1) and human cervix carcinoma cells (HeLa)] were verified by STR analysis (BMR Genomics, Italy).
6
Estrogen Signaling in Breast Cancer
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Cell culture and reagents.
17β-estradiol, DMEM (with and without phenol red) and fetal calf serum were purchased from Sigma-Aldrich (St. Louis, MO). Bradford protein assay kit as well as anti-mouse and antirabbit secondary antibodies were obtained from Bio-Rad (Hercules, CA). Antibodies against ERα (F-10 mouse), cyclin D1 (H-295 rabbit), cathepsin D (H75 rabbit), pS2 (FL-84 rabbit), were obtained from Santa Cruz Biotechnology (Santa Cruz, CA); anti-vinculin antibody was from Sigma-Aldrich (St. Louis, MO). Chemiluminescence reagent for Western blotting was obtained from BioRad Laboratories (Hercules, CA, USA). Faslodex (i.e., fulvestrant or ICI182,780) and 4OH-tamoxifen were purchased by Tocris (USA). FDA-approved drug library was purchased by Selleck Chemicals (USA). All the other products were from Sigma-Aldrich. Analytical-or reagentgrade products were used without further purification. The identities of all of the used cell lines [i.e., human breast carcinoma cells (MCF-7; ZR-75-1)] were verified by STR analysis (BMR Genomics, Italy).
17β-estradiol, DMEM (with and without phenol red) and fetal calf serum were purchased from Sigma-Aldrich (St. Louis, MO). Bradford protein assay kit as well as anti-mouse and antirabbit secondary antibodies were obtained from Bio-Rad (Hercules, CA). Antibodies against ERα (F-10 mouse), cyclin D1 (H-295 rabbit), cathepsin D (H75 rabbit), pS2 (FL-84 rabbit), were obtained from Santa Cruz Biotechnology (Santa Cruz, CA); anti-vinculin antibody was from Sigma-Aldrich (St. Louis, MO). Chemiluminescence reagent for Western blotting was obtained from BioRad Laboratories (Hercules, CA, USA). Faslodex (i.e., fulvestrant or ICI182,780) and 4OH-tamoxifen were purchased by Tocris (USA). FDA-approved drug library was purchased by Selleck Chemicals (USA). All the other products were from Sigma-Aldrich. Analytical-or reagentgrade products were used without further purification. The identities of all of the used cell lines [i.e., human breast carcinoma cells (MCF-7; ZR-75-1)] were verified by STR analysis (BMR Genomics, Italy).
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