Isolation:Article Title: Metabolic Adaptations of Prostate Cancer Cells Under Chronic Fructose Stimulation
Article Snippet: .. A fraction enriched in mitochondria was isolated from PC-3 and LNCaP cell lysates obtained with MSH buffer (230 mM mannitol, 70 mM sucrose, 5 mM Hepes, pH 7.4) supplemented with phosphatase (NaF 20x, Na2P2O7 300x, Na3VO4 100x) and protease (#78429 Thermo Fisher Scientific) inhibitors. ..
Saline:Article Title: Identifying a novel Mecp2-mediated epigenetic mechanism controlling Lonp1 in the hippocampus and its disruption by aging
Article Snippet: .. The following reagents and antibodies were used: Cycloheximide (CAS 66–81-9, Sigma Aldrich), Chloroquine diphosphate salt (C6628, Sigma Aldrich), MG132 (ab141003, Abcam), MSH buffer (230 mM Mannitol, 70 mM sucrose, 5 mM HEPES, 1 mM EDTA, pH = 7.4) supplemented with phosphatase (NaF 25 mM, Na 2 P 2 O 7 30 μM, Na 3 VO 4 100 mM) and protease (catalog number 78429, Thermo Fisher Scientific) inhibitors, Saline Sodium Citrate (SSC) Buffer (3 M NaCl, 0,3 M Sodium Citrate, pH = 7.0), DNA Denaturing Buffer (200 mM NaOH, 20 mM EDTA), BCA Protein Assay Kit (23,227, Thermo Fisher Scientific). .. The primary antibodies used were mouse anti-Actin (1:1000, sc-1616, Santa Cruz Biotechnology, Inc), rabbit anti-VDAC (1:1000, sc-390996, Santa Cruz Biotechnology, Inc), rabbit anti-GAPDH (1:1000, sc-25778, Santa Cruz Biotechnology, Inc.), rabbit anti-Lamin A (1:1000, sc-20680, Santa Cruz Biotechnology, Inc.), rabbit anti-Lamin B1 (1:1000, sc-365962, Santa Cruz Biotechnology, Inc.), rabbit anti-Lonp1 (1:1000, PA551692 , Termofisher Scientific), rabbit anti-Mecp2 (ab2828, Abcam), rabbit anti-Mecp2 (1:1000, 07–013, Merck Millipore), rabbit anti-Mecp2pSer80 (1:1000, AB2793344, Active Motif), rabbit anti-Mecp2 pSer421 (1:1000, AB254050 , Active Motif), rabbit anti-5-Methylcitosine (5-mC) (1:1000, D3S2Z, Cell signaling Technology), mouse anti-β catenin (1:1000, sc-133240, Santa Cruz Biotechnology, Inc.), rabbit anti-LC3B (D11) (1:1000, 3868, cell signaling Technology), rabbit anti-SQSMT1-p62 (1:1000, 5114, cell signaling Technology).
Article Title: Identifying a novel Mecp2-mediated epigenetic mechanism controlling Lonp1 in the hippocampus and its disruption by aging.
Article Snippet: .. The following reagents and antibodies were used: Cycloheximide (CAS 66–81-9, Sigma Aldrich), Chloroquine diphosphate salt (C6628, Sigma Aldrich), MG132 (ab141003, Abcam), MSH buffer (230 mM Mannitol, 70 mM sucrose, 5 mM HEPES, 1 mM EDTA, pH = 7.4) supplemented with phosphatase (NaF 25 mM, Na2P2O7 30 μM, Na3VO4 100 mM) and protease (catalog number 78429, Thermo Fisher Scientific) inhibitors, Saline Sodium Citrate (SSC) Buffer (3 M NaCl, 0,3 M Sodium Citrate, pH = 7.0), DNA Denaturing Buffer (200 mM NaOH, 20 mM EDTA), BCA Protein Assay Kit (23,227, Thermo Fisher Scientific). .. The primary antibodies used were mouse anti-Actin (1:1000, sc-1616, Santa Cruz Biotechnology, Inc), rabbit anti-VDAC (1:1000, sc-390996, Santa Cruz Biotechnology, Inc), rabbit anti-GAPDH (1:1000, sc-25778, Santa Cruz Biotechnology, Inc.), rabbit antiLamin A (1:1000, sc-20680, Santa Cruz Biotechnology, Inc.), rabbit anti-Lamin B1 (1:1000, sc-365962, Santa Cruz Biotechnology, Inc.), rabbit anti-Lonp1 (1:1000, PA551692, Termofisher Scientific), rabbit anti-Mecp2 (ab2828, Abcam), rabbit anti-Mecp2 (1:1000, 07–013, Merck Millipore), rabbit anti-Mecp2pSer80 (1:1000, AB2793344, Active Motif), rabbit anti-Mecp2 pSer421 (1:1000, AB254050, Active Motif), rabbit anti-5Methylcitosine (5-mC) (1:1000, D3S2Z, Cell signaling Technology), mouse anti-β catenin (1:1000, sc-133240, Santa Cruz Biotechnology, Inc.), rabbit anti-LC3B (D11) (1:1000, 3868, cell signaling Technology), rabbit antiSQSMT1-p62 (1:1000, 5114, cell signaling Technology).
Bicinchoninic Acid Protein Assay:Article Title: Identifying a novel Mecp2-mediated epigenetic mechanism controlling Lonp1 in the hippocampus and its disruption by aging
Article Snippet: .. The following reagents and antibodies were used: Cycloheximide (CAS 66–81-9, Sigma Aldrich), Chloroquine diphosphate salt (C6628, Sigma Aldrich), MG132 (ab141003, Abcam), MSH buffer (230 mM Mannitol, 70 mM sucrose, 5 mM HEPES, 1 mM EDTA, pH = 7.4) supplemented with phosphatase (NaF 25 mM, Na 2 P 2 O 7 30 μM, Na 3 VO 4 100 mM) and protease (catalog number 78429, Thermo Fisher Scientific) inhibitors, Saline Sodium Citrate (SSC) Buffer (3 M NaCl, 0,3 M Sodium Citrate, pH = 7.0), DNA Denaturing Buffer (200 mM NaOH, 20 mM EDTA), BCA Protein Assay Kit (23,227, Thermo Fisher Scientific). .. The primary antibodies used were mouse anti-Actin (1:1000, sc-1616, Santa Cruz Biotechnology, Inc), rabbit anti-VDAC (1:1000, sc-390996, Santa Cruz Biotechnology, Inc), rabbit anti-GAPDH (1:1000, sc-25778, Santa Cruz Biotechnology, Inc.), rabbit anti-Lamin A (1:1000, sc-20680, Santa Cruz Biotechnology, Inc.), rabbit anti-Lamin B1 (1:1000, sc-365962, Santa Cruz Biotechnology, Inc.), rabbit anti-Lonp1 (1:1000, PA551692 , Termofisher Scientific), rabbit anti-Mecp2 (ab2828, Abcam), rabbit anti-Mecp2 (1:1000, 07–013, Merck Millipore), rabbit anti-Mecp2pSer80 (1:1000, AB2793344, Active Motif), rabbit anti-Mecp2 pSer421 (1:1000, AB254050 , Active Motif), rabbit anti-5-Methylcitosine (5-mC) (1:1000, D3S2Z, Cell signaling Technology), mouse anti-β catenin (1:1000, sc-133240, Santa Cruz Biotechnology, Inc.), rabbit anti-LC3B (D11) (1:1000, 3868, cell signaling Technology), rabbit anti-SQSMT1-p62 (1:1000, 5114, cell signaling Technology).
Article Title: Identifying a novel Mecp2-mediated epigenetic mechanism controlling Lonp1 in the hippocampus and its disruption by aging.
Article Snippet: .. The following reagents and antibodies were used: Cycloheximide (CAS 66–81-9, Sigma Aldrich), Chloroquine diphosphate salt (C6628, Sigma Aldrich), MG132 (ab141003, Abcam), MSH buffer (230 mM Mannitol, 70 mM sucrose, 5 mM HEPES, 1 mM EDTA, pH = 7.4) supplemented with phosphatase (NaF 25 mM, Na2P2O7 30 μM, Na3VO4 100 mM) and protease (catalog number 78429, Thermo Fisher Scientific) inhibitors, Saline Sodium Citrate (SSC) Buffer (3 M NaCl, 0,3 M Sodium Citrate, pH = 7.0), DNA Denaturing Buffer (200 mM NaOH, 20 mM EDTA), BCA Protein Assay Kit (23,227, Thermo Fisher Scientific). .. The primary antibodies used were mouse anti-Actin (1:1000, sc-1616, Santa Cruz Biotechnology, Inc), rabbit anti-VDAC (1:1000, sc-390996, Santa Cruz Biotechnology, Inc), rabbit anti-GAPDH (1:1000, sc-25778, Santa Cruz Biotechnology, Inc.), rabbit antiLamin A (1:1000, sc-20680, Santa Cruz Biotechnology, Inc.), rabbit anti-Lamin B1 (1:1000, sc-365962, Santa Cruz Biotechnology, Inc.), rabbit anti-Lonp1 (1:1000, PA551692, Termofisher Scientific), rabbit anti-Mecp2 (ab2828, Abcam), rabbit anti-Mecp2 (1:1000, 07–013, Merck Millipore), rabbit anti-Mecp2pSer80 (1:1000, AB2793344, Active Motif), rabbit anti-Mecp2 pSer421 (1:1000, AB254050, Active Motif), rabbit anti-5Methylcitosine (5-mC) (1:1000, D3S2Z, Cell signaling Technology), mouse anti-β catenin (1:1000, sc-133240, Santa Cruz Biotechnology, Inc.), rabbit anti-LC3B (D11) (1:1000, 3868, cell signaling Technology), rabbit antiSQSMT1-p62 (1:1000, 5114, cell signaling Technology).
Protease Inhibitor:Article Title: Lonp1 Inhibition Disrupts Mitochondrial Function in the hippocampus and Drives an Aging-Like Synaptic and Cognitive Phenotype in adult SAMP8 mice.
Article Snippet: Lonp1 is the main mitochondrial matrix protease responsible for maintaining mitochondrial proteostasis through the degradation of damaged or misfolded proteins.. Although impaired Lonp1 expression or activity has been linked to mitochondrial dysfunction and oxidative stress in peripheral tissues and non-neuronal cells, its role in the brain, and particularly in hippocampal function, remains unexplored.. Here, we provide the first in vivo evidence that Lonp1 activity is a critical regulator of mitochondrial redox homeostasis, synaptic integrity, and learning in the hippocampus.
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