To analyze PTHrP secretion from ER+ tumor cells and its E2 dependency, ER+ MCF-7 cells or ER+ tumor cells isolated from MCF-7 BMET were plated in 24-well plates at a density of 1.3 × 105 cells/well in E2-depleted media [phenol red-free DMEM (Invitrogen), 10% charcoal-stripped FBS (Valley Biomedical, Winchester, VA), 1% penicillin/streptomycin (Thermo Fisher), and 200 mM L-Glutamine (Sigma Aldrich, St. Louis, MO)] for 4 days, during which time cell number did not change for any cell line (data not shown), prior to treatment with E2 (10−11-10−6 M, as indicated; Sigma Aldrich), an ERα specific agonist propyl pyrazole triol (PPT; 10−8 M; Tocris, Minneapolis, MN), an ERα specific antagonist methyl-piperidinopyrazole hydrate (MPP; 10−6 M, Tocris), or vehicle control for 48 or 52 h, as indicated. Conditioned media, stored at −80 °C after addition of protease inhibitors (Sigma Aldrich), were assayed for secreted PTHrP using a commercial immunoradiometric assay (Beckman Coulter, Brea, CA). A lack of treatment effect on cell number during the 48 or 52-h incubation was verified using a commercial MTT assay (ATCC).
Methyl piperidinopyrazole hydrate mpp
Manufactured by Bio-Techne
Methyl-piperidinopyrazole hydrate (MPP) is a chemical compound that functions as a selective antagonist for the histamine H3 receptor. It is commonly used as a research tool in the study of the histaminergic system and its potential therapeutic applications.
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Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Protease inhibitor, by Merck Group (2 mentions)
Mtt assay, by American Type Culture Collection (2 mentions)
L glutamine, by Merck Group (2 mentions)
Phenol red free dmem, by Thermo Fisher Scientific (2 mentions)
Charcoal stripped fbs, by Valley Biomedical (2 mentions)
Propyl pyrazole triol ppt, by Bio-Techne (2 mentions)
2 protocols using methyl piperidinopyrazole hydrate mpp
1
Estrogen-Dependent PTHrP Secretion in ER+ Tumor Cells
2
Estrogen-Dependent PTHrP Secretion in ER+ Tumor Cells
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To analyze PTHrP secretion from ER+ tumor cells and its E2 dependency, ER+ MCF-7 cells or ER+ tumor cells isolated from MCF-7 BMET were plated in 24-well plates at a density of 1.3 × 105 cells/well in E2-depleted media [phenol red-free DMEM (Invitrogen), 10% charcoal-stripped FBS (Valley Biomedical, Winchester, VA), 1% penicillin/streptomycin (Thermo Fisher), and 200 mM L-Glutamine (Sigma Aldrich, St. Louis, MO)] for 4 days, during which time cell number did not change for any cell line (data not shown), prior to treatment with E2 (10−11-10−6 M, as indicated; Sigma Aldrich), an ERα specific agonist propyl pyrazole triol (PPT; 10−8 M; Tocris, Minneapolis, MN), an ERα specific antagonist methyl-piperidinopyrazole hydrate (MPP; 10−6 M, Tocris), or vehicle control for 48 or 52 h, as indicated. Conditioned media, stored at −80 °C after addition of protease inhibitors (Sigma Aldrich), were assayed for secreted PTHrP using a commercial immunoradiometric assay (Beckman Coulter, Brea, CA). A lack of treatment effect on cell number during the 48 or 52-h incubation was verified using a commercial MTT assay (ATCC).
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