Incubation:Article Title: The cognitive defects of neonatally irradiated mice are accompanied by changed synaptic plasticity, adult neurogenesis and neuroinflammation.
Article Snippet: .. Hippocampal and cortical protein extracts (15 μg) were separated on 12% SDS polyacrylamide gels and were blotted to nitrocellulose membranes (GE Healthcare, Germany) via BIO-RAD criterionTM Blotter system at 100 V for 2 h. Membranes were blocked with RotiR-Block solution (Roth, Germany), washed and incubated overnight at 4°C with primary antibody dilutions as indicated by the manufacturer (GAPDH – sc-47724 Santa Cruz, Germany; CDC42 – ab106374 Abcam, Germany; Rac1 – ab33186 Abcam, Germany; p-PAK1 – 2601 Cell Signaling, Germany; Fascin – ab74487 Abcam, Germany; Cofilin – 3312 Cell Signaling, Germany; Rho-GDIα – ab108977 Epitomics, Germany; p-LIMK1/2 – sc-28409-R Santa Cruz, Germany; p-Cofilin – 3311 Cell Signalling, Germany; LIMK1 – 3842 Cell Signaling, Germany; p-Rho-GDIα – sc-33047 Santa Cruz, Germany; TNFα – 3707 Cell Signaling, Germany; p-IGF1Rβ/INSRβ – 3021 Cell Signalling, Germany; Arc – ab118929 Abcam, Germany; c-Fos – sc52 Santa Cruz, Germany; p-CREB – 9191 Cell Signaling, Germany; CREB – 4820 Cell Signaling, Germany). .. After a washing step, blots were incubated with appropriate horseradish peroxidase-conjugated secondary antibodies in 8% milk for 1 h at room temperature and developed using the ECL system (GE Healthcare, Germany) using standard procedures.
Article Title: Age-related effects of X-ray irradiation on mouse hippocampus
Article Snippet: .. Proteins separated by 4-12% SDS-PAGE were transferred to nitrocellulose membranes (GE Healthcare) using BIO-RAD CriterionTM Blotter system at 100 V for 2 h. The membranes were blocked using 8% milk in PBS, pH 7.4, for 2 h at room temperature, washed three times in 10 mM Tris-HCl, pH 7.4, 150 mM NaCl for 5 min and incubated overnight at 4°C with primary antibodies using dilutions recommended by the manufacturer. .. Immunoblot analysis of hippocampus protein lysate was performed using anti- postsynaptic protein PSD-95 (Dlg4) (ABIN1742271, Atlanta, GA, USA), anti-Rac1 (ab33186, abcam), anti-Hspd1 (ab46798, abcam), anti-Sod1 (sc-11407, Santa Cruz Biotechnology, Dallas, Texas USA), and anti-tubulin alpha (GTX72360, Biocompare, San Francisco, CA, USA).
Article Title: Ionising radiation immediately impairs synaptic plasticity-associated cytoskeletal signalling pathways in HT22 cells and in mouse brain: an in vitro/in vivo comparison study.
Article Snippet: .. Quantification of Rac1, cofilin and phospho-cofilin expression levels via immunoblotting Protein extracts of cells and brain tissues (15 mg) were separated on 12% SDS polyacrylamide gels and transferred to nitrocellulose membranes (GE Healthcare) via BIO-RAD CriterionTM Blotter system at 100 V for 2 h. Membranes were blocked with RotiRBlock solution (Roth), washed and incubated overnight at 4uC with primary antibody dilutions as recommended by the manufacturer (GAPDH – sc-47724 [murine monoclonal IgG1 raised against recombinant GAPDH of human origin; Santa Cruz], Rac1– ab33186 [murine monoclonal IgG2b raised against full-length recombinant Rac1 of human origin; Abcam], cofilin –3312 [rabbit polyclonal antibody produced by immunising rabbits with a synthetic peptide corresponding to residues surrounding Ser3 of human cofilin origin; Cell Signalling], p-Cofilin (Ser3) –3311 [rabbit polyclonal antibody by immunising animals with a synthetic phospho-peptide corresponding to residues surrounding Ser3 of human cofilin; Cell Signalling]). .. After a washing step, blots were incubated with appropriate horseradish peroxidase-conjugated secondary antibody in 8% milk for 1 h at room temperature and developed using ECL system (GE Healthcare) using standard protocol from the manufacturer.
Article Title: Ionising Radiation Immediately Impairs Synaptic Plasticity-Associated Cytoskeletal Signalling Pathways in HT22 Cells and in Mouse Brain: An In Vitro/In Vivo Comparison Study
Article Snippet: .. Protein extracts of cells and brain tissues (15 μg) were separated on 12% SDS polyacrylamide gels and transferred to nitrocellulose membranes (GE Healthcare) via BIO-RAD CriterionTM Blotter system at 100 V for 2 h. Membranes were blocked with Roti R -Block solution (Roth), washed and incubated overnight at 4°C with primary antibody dilutions as recommended by the manufacturer (GAPDH – sc-47724 [murine monoclonal IgG1 raised against recombinant GAPDH of human origin; Santa Cruz], Rac1– ab33186 [murine monoclonal IgG2b raised against full-length recombinant Rac1 of human origin; Abcam], cofilin –3312 [rabbit polyclonal antibody produced by immunising rabbits with a synthetic peptide corresponding to residues surrounding Ser3 of human cofilin origin; Cell Signalling], p-Cofilin (Ser3) –3311 [rabbit polyclonal antibody by immunising animals with a synthetic phospho-peptide corresponding to residues surrounding Ser3 of human cofilin; Cell Signalling]). .. After a washing step, blots were incubated with appropriate horseradish peroxidase-conjugated secondary antibody in 8% milk for 1 h at room temperature and developed using ECL system (GE Healthcare) using standard protocol from the manufacturer.
Article Title: The cognitive defects of neonatally irradiated mice are accompanied by changed synaptic plasticity, adult neurogenesis and neuroinflammation
Article Snippet: .. Hippocampal and cortical protein extracts (15 μg) were separated on 12% SDS polyacrylamide gels and were blotted to nitrocellulose membranes (GE Healthcare, Germany) via BIO-RAD criterionTM Blotter system at 100 V for 2 h. Membranes were blocked with Roti R -Block solution (Roth, Germany), washed and incubated overnight at 4°C with primary antibody dilutions as indicated by the manufacturer (GAPDH – sc-47724 Santa Cruz, Germany; CDC42 – ab106374 Abcam, Germany; Rac1 – ab33186 Abcam, Germany; p-PAK1 – 2601 Cell Signaling, Germany; Fascin – ab74487 Abcam, Germany; Cofilin – 3312 Cell Signaling, Germany; Rho-GDIα – ab108977 Epitomics, Germany; p-LIMK1/2 – sc-28409-R Santa Cruz, Germany; p-Cofilin – 3311 Cell Signalling, Germany; LIMK1 – 3842 Cell Signaling, Germany; p-Rho-GDIα – sc-33047 Santa Cruz, Germany; TNFα – 3707 Cell Signaling, Germany; p-IGF1Rβ/INSRβ – 3021 Cell Signalling, Germany; Arc – ab118929 Abcam, Germany; c-Fos – sc-52 Santa Cruz, Germany; p-CREB – 9191 Cell Signaling, Germany; CREB – 4820 Cell Signaling, Germany). .. After a washing step, blots were incubated with appropriate horseradish peroxidase-conjugated secondary antibodies in 8% milk for 1 h at room temperature and developed using the ECL system (GE Healthcare, Germany) using standard procedures.
SDS Page:Article Title: Age-related effects of X-ray irradiation on mouse hippocampus
Article Snippet: .. Proteins separated by 4-12% SDS-PAGE were transferred to nitrocellulose membranes (GE Healthcare) using BIO-RAD CriterionTM Blotter system at 100 V for 2 h. The membranes were blocked using 8% milk in PBS, pH 7.4, for 2 h at room temperature, washed three times in 10 mM Tris-HCl, pH 7.4, 150 mM NaCl for 5 min and incubated overnight at 4°C with primary antibodies using dilutions recommended by the manufacturer. .. Immunoblot analysis of hippocampus protein lysate was performed using anti- postsynaptic protein PSD-95 (Dlg4) (ABIN1742271, Atlanta, GA, USA), anti-Rac1 (ab33186, abcam), anti-Hspd1 (ab46798, abcam), anti-Sod1 (sc-11407, Santa Cruz Biotechnology, Dallas, Texas USA), and anti-tubulin alpha (GTX72360, Biocompare, San Francisco, CA, USA).
Expressing:Article Title: Ionising radiation immediately impairs synaptic plasticity-associated cytoskeletal signalling pathways in HT22 cells and in mouse brain: an in vitro/in vivo comparison study.
Article Snippet: .. Quantification of Rac1, cofilin and phospho-cofilin expression levels via immunoblotting Protein extracts of cells and brain tissues (15 mg) were separated on 12% SDS polyacrylamide gels and transferred to nitrocellulose membranes (GE Healthcare) via BIO-RAD CriterionTM Blotter system at 100 V for 2 h. Membranes were blocked with RotiRBlock solution (Roth), washed and incubated overnight at 4uC with primary antibody dilutions as recommended by the manufacturer (GAPDH – sc-47724 [murine monoclonal IgG1 raised against recombinant GAPDH of human origin; Santa Cruz], Rac1– ab33186 [murine monoclonal IgG2b raised against full-length recombinant Rac1 of human origin; Abcam], cofilin –3312 [rabbit polyclonal antibody produced by immunising rabbits with a synthetic peptide corresponding to residues surrounding Ser3 of human cofilin origin; Cell Signalling], p-Cofilin (Ser3) –3311 [rabbit polyclonal antibody by immunising animals with a synthetic phospho-peptide corresponding to residues surrounding Ser3 of human cofilin; Cell Signalling]). .. After a washing step, blots were incubated with appropriate horseradish peroxidase-conjugated secondary antibody in 8% milk for 1 h at room temperature and developed using ECL system (GE Healthcare) using standard protocol from the manufacturer.
Western Blot:Article Title: Ionising radiation immediately impairs synaptic plasticity-associated cytoskeletal signalling pathways in HT22 cells and in mouse brain: an in vitro/in vivo comparison study.
Article Snippet: .. Quantification of Rac1, cofilin and phospho-cofilin expression levels via immunoblotting Protein extracts of cells and brain tissues (15 mg) were separated on 12% SDS polyacrylamide gels and transferred to nitrocellulose membranes (GE Healthcare) via BIO-RAD CriterionTM Blotter system at 100 V for 2 h. Membranes were blocked with RotiRBlock solution (Roth), washed and incubated overnight at 4uC with primary antibody dilutions as recommended by the manufacturer (GAPDH – sc-47724 [murine monoclonal IgG1 raised against recombinant GAPDH of human origin; Santa Cruz], Rac1– ab33186 [murine monoclonal IgG2b raised against full-length recombinant Rac1 of human origin; Abcam], cofilin –3312 [rabbit polyclonal antibody produced by immunising rabbits with a synthetic peptide corresponding to residues surrounding Ser3 of human cofilin origin; Cell Signalling], p-Cofilin (Ser3) –3311 [rabbit polyclonal antibody by immunising animals with a synthetic phospho-peptide corresponding to residues surrounding Ser3 of human cofilin; Cell Signalling]). .. After a washing step, blots were incubated with appropriate horseradish peroxidase-conjugated secondary antibody in 8% milk for 1 h at room temperature and developed using ECL system (GE Healthcare) using standard protocol from the manufacturer.
Recombinant:Article Title: Ionising radiation immediately impairs synaptic plasticity-associated cytoskeletal signalling pathways in HT22 cells and in mouse brain: an in vitro/in vivo comparison study.
Article Snippet: .. Quantification of Rac1, cofilin and phospho-cofilin expression levels via immunoblotting Protein extracts of cells and brain tissues (15 mg) were separated on 12% SDS polyacrylamide gels and transferred to nitrocellulose membranes (GE Healthcare) via BIO-RAD CriterionTM Blotter system at 100 V for 2 h. Membranes were blocked with RotiRBlock solution (Roth), washed and incubated overnight at 4uC with primary antibody dilutions as recommended by the manufacturer (GAPDH – sc-47724 [murine monoclonal IgG1 raised against recombinant GAPDH of human origin; Santa Cruz], Rac1– ab33186 [murine monoclonal IgG2b raised against full-length recombinant Rac1 of human origin; Abcam], cofilin –3312 [rabbit polyclonal antibody produced by immunising rabbits with a synthetic peptide corresponding to residues surrounding Ser3 of human cofilin origin; Cell Signalling], p-Cofilin (Ser3) –3311 [rabbit polyclonal antibody by immunising animals with a synthetic phospho-peptide corresponding to residues surrounding Ser3 of human cofilin; Cell Signalling]). .. After a washing step, blots were incubated with appropriate horseradish peroxidase-conjugated secondary antibody in 8% milk for 1 h at room temperature and developed using ECL system (GE Healthcare) using standard protocol from the manufacturer.
Article Title: Ionising Radiation Immediately Impairs Synaptic Plasticity-Associated Cytoskeletal Signalling Pathways in HT22 Cells and in Mouse Brain: An In Vitro/In Vivo Comparison Study
Article Snippet: .. Protein extracts of cells and brain tissues (15 μg) were separated on 12% SDS polyacrylamide gels and transferred to nitrocellulose membranes (GE Healthcare) via BIO-RAD CriterionTM Blotter system at 100 V for 2 h. Membranes were blocked with Roti R -Block solution (Roth), washed and incubated overnight at 4°C with primary antibody dilutions as recommended by the manufacturer (GAPDH – sc-47724 [murine monoclonal IgG1 raised against recombinant GAPDH of human origin; Santa Cruz], Rac1– ab33186 [murine monoclonal IgG2b raised against full-length recombinant Rac1 of human origin; Abcam], cofilin –3312 [rabbit polyclonal antibody produced by immunising rabbits with a synthetic peptide corresponding to residues surrounding Ser3 of human cofilin origin; Cell Signalling], p-Cofilin (Ser3) –3311 [rabbit polyclonal antibody by immunising animals with a synthetic phospho-peptide corresponding to residues surrounding Ser3 of human cofilin; Cell Signalling]). .. After a washing step, blots were incubated with appropriate horseradish peroxidase-conjugated secondary antibody in 8% milk for 1 h at room temperature and developed using ECL system (GE Healthcare) using standard protocol from the manufacturer.
Produced:Article Title: Ionising radiation immediately impairs synaptic plasticity-associated cytoskeletal signalling pathways in HT22 cells and in mouse brain: an in vitro/in vivo comparison study.
Article Snippet: .. Quantification of Rac1, cofilin and phospho-cofilin expression levels via immunoblotting Protein extracts of cells and brain tissues (15 mg) were separated on 12% SDS polyacrylamide gels and transferred to nitrocellulose membranes (GE Healthcare) via BIO-RAD CriterionTM Blotter system at 100 V for 2 h. Membranes were blocked with RotiRBlock solution (Roth), washed and incubated overnight at 4uC with primary antibody dilutions as recommended by the manufacturer (GAPDH – sc-47724 [murine monoclonal IgG1 raised against recombinant GAPDH of human origin; Santa Cruz], Rac1– ab33186 [murine monoclonal IgG2b raised against full-length recombinant Rac1 of human origin; Abcam], cofilin –3312 [rabbit polyclonal antibody produced by immunising rabbits with a synthetic peptide corresponding to residues surrounding Ser3 of human cofilin origin; Cell Signalling], p-Cofilin (Ser3) –3311 [rabbit polyclonal antibody by immunising animals with a synthetic phospho-peptide corresponding to residues surrounding Ser3 of human cofilin; Cell Signalling]). .. After a washing step, blots were incubated with appropriate horseradish peroxidase-conjugated secondary antibody in 8% milk for 1 h at room temperature and developed using ECL system (GE Healthcare) using standard protocol from the manufacturer.
Article Title: Ionising Radiation Immediately Impairs Synaptic Plasticity-Associated Cytoskeletal Signalling Pathways in HT22 Cells and in Mouse Brain: An In Vitro/In Vivo Comparison Study
Article Snippet: .. Protein extracts of cells and brain tissues (15 μg) were separated on 12% SDS polyacrylamide gels and transferred to nitrocellulose membranes (GE Healthcare) via BIO-RAD CriterionTM Blotter system at 100 V for 2 h. Membranes were blocked with Roti R -Block solution (Roth), washed and incubated overnight at 4°C with primary antibody dilutions as recommended by the manufacturer (GAPDH – sc-47724 [murine monoclonal IgG1 raised against recombinant GAPDH of human origin; Santa Cruz], Rac1– ab33186 [murine monoclonal IgG2b raised against full-length recombinant Rac1 of human origin; Abcam], cofilin –3312 [rabbit polyclonal antibody produced by immunising rabbits with a synthetic peptide corresponding to residues surrounding Ser3 of human cofilin origin; Cell Signalling], p-Cofilin (Ser3) –3311 [rabbit polyclonal antibody by immunising animals with a synthetic phospho-peptide corresponding to residues surrounding Ser3 of human cofilin; Cell Signalling]). .. After a washing step, blots were incubated with appropriate horseradish peroxidase-conjugated secondary antibody in 8% milk for 1 h at room temperature and developed using ECL system (GE Healthcare) using standard protocol from the manufacturer.
Blocking Assay:Article Title: Ionising Radiation Immediately Impairs Synaptic Plasticity-Associated Cytoskeletal Signalling Pathways in HT22 Cells and in Mouse Brain: An In Vitro/In Vivo Comparison Study
Article Snippet: .. Protein extracts of cells and brain tissues (15 μg) were separated on 12% SDS polyacrylamide gels and transferred to nitrocellulose membranes (GE Healthcare) via BIO-RAD CriterionTM Blotter system at 100 V for 2 h. Membranes were blocked with Roti R -Block solution (Roth), washed and incubated overnight at 4°C with primary antibody dilutions as recommended by the manufacturer (GAPDH – sc-47724 [murine monoclonal IgG1 raised against recombinant GAPDH of human origin; Santa Cruz], Rac1– ab33186 [murine monoclonal IgG2b raised against full-length recombinant Rac1 of human origin; Abcam], cofilin –3312 [rabbit polyclonal antibody produced by immunising rabbits with a synthetic peptide corresponding to residues surrounding Ser3 of human cofilin origin; Cell Signalling], p-Cofilin (Ser3) –3311 [rabbit polyclonal antibody by immunising animals with a synthetic phospho-peptide corresponding to residues surrounding Ser3 of human cofilin; Cell Signalling]). .. After a washing step, blots were incubated with appropriate horseradish peroxidase-conjugated secondary antibody in 8% milk for 1 h at room temperature and developed using ECL system (GE Healthcare) using standard protocol from the manufacturer.
Article Title: The cognitive defects of neonatally irradiated mice are accompanied by changed synaptic plasticity, adult neurogenesis and neuroinflammation
Article Snippet: .. Hippocampal and cortical protein extracts (15 μg) were separated on 12% SDS polyacrylamide gels and were blotted to nitrocellulose membranes (GE Healthcare, Germany) via BIO-RAD criterionTM Blotter system at 100 V for 2 h. Membranes were blocked with Roti R -Block solution (Roth, Germany), washed and incubated overnight at 4°C with primary antibody dilutions as indicated by the manufacturer (GAPDH – sc-47724 Santa Cruz, Germany; CDC42 – ab106374 Abcam, Germany; Rac1 – ab33186 Abcam, Germany; p-PAK1 – 2601 Cell Signaling, Germany; Fascin – ab74487 Abcam, Germany; Cofilin – 3312 Cell Signaling, Germany; Rho-GDIα – ab108977 Epitomics, Germany; p-LIMK1/2 – sc-28409-R Santa Cruz, Germany; p-Cofilin – 3311 Cell Signalling, Germany; LIMK1 – 3842 Cell Signaling, Germany; p-Rho-GDIα – sc-33047 Santa Cruz, Germany; TNFα – 3707 Cell Signaling, Germany; p-IGF1Rβ/INSRβ – 3021 Cell Signalling, Germany; Arc – ab118929 Abcam, Germany; c-Fos – sc-52 Santa Cruz, Germany; p-CREB – 9191 Cell Signaling, Germany; CREB – 4820 Cell Signaling, Germany). .. After a washing step, blots were incubated with appropriate horseradish peroxidase-conjugated secondary antibodies in 8% milk for 1 h at room temperature and developed using the ECL system (GE Healthcare, Germany) using standard procedures.
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