Saline:Article Title: A preclinical and phase I clinical study of ex vivo-expanded amyloid beta-specific human regulatory T cells in Alzheimer's disease.
Article Snippet: .. Each blot was blocked for 30 min in Tris-buffered saline/Tween-20 (10 mM Tris base, 150 mM NaCl, 0.05 % Tween-20) containing 5 % fat-free milk and incubated with the following primary antibodies: CD86 (1:1000 dilution, Cell Signaling Technology), NOS2 (1:2000 dilution, Santa Cruz Biotechnology), TREM2 (1:1000 dilution, MAB1828 R&D Systems), or β-actin (1:5000 dilution, Santa Cruz Biotechnology), overnight at 4°C. ..
Article Title: H2AX phosphorylation driven by the SET-PP2A axis maintains glioma stem cell properties
Article Snippet: .. The membrane was blocked with 5% nonfat 174 milk in Tris-buffered saline-Tween 20 and incubated overnight at 4 °C with the following 175 primary antibodies: Rabbit anti-SOX2 (CST, Cat. #3579S, 1:1,000), Rabbit anti-OLIG2 176 (CST, Cat. #2748, 1:1000), Rabbit anti-NANOG (CST, Cat. #4903, 1:1,000), Mouse anti-177 GFAP (BioLegend, Cat. #644702, 1:1,000), Rabbit anti-H2AX (CST, Cat. #7631, 178 1:1,000), mouse anti-γH2AX (Millipore Sigma, Cat. #05-636, 1: 2,000), Rabbit anti-179 Tubulin (CST, Cat. #2144, 1:1,000), Rabbit anti-GAPDH (CST, Cat. #2118, 1:1,000), 180 Rabbit anti-ATM (CST, Cat. #2873, 1:1,000), Rabbit anti-pATM (S1981) (CST, Cat. 181 #5883, 1:1,000), Rabbit anti-ATR (CST, Cat. #13934, 1:1000), Rabbit anti-pATR (S428) 182 Jo rna l P repro of 9 (CST, Cat. #2853, 1:1000), Rabbit anti-pRPA32 (S33) (CST, Cat. #10148, 1:1,000), 183 Rabbit anti-RPA32 (CST, Cat. #35869, 1:1,000), Rabbit anti-SET/TAF1 antibody 184 (ABclonal, Cat. #A9173, 1:500), mouse anti-I2PP2A (SET) antibody (Santa Cruz 185 Biotechnology, Cat. #sc-133138, 1:500). .. Following multiple washes, the membrane was 186 incubated for 1 h at room temperature with HRP-conjugated goat anti-rabbit or goat anti-187 mouse secondary antibodies (Millipore Sigma, Cat. #12-348, 12-349).
Article Title: Regulation of skeletal muscle development and metabolism in broiler chickens by Urolithin A through threonine kinase 1 pathway activation
Article Snippet: .. After blocking the membrane with 5% (wt/vol) skim milk in Tris-buffered saline/Tween 20 for 1 h at 37°C, incubation was carried out overnight at 4°C with GAPDH (Proteintech, Wuhan, China), Akt1 (Cell Signaling Technology, Massachusetts, USA; ), and p-Akt1 (Cell Signaling Technology, Massachusetts, USA) antibodies. .. Subsequently, incubation with a peroxidase-conjugated anti-rabbit IgG antibody (Proteintech, Wuhan, China) was performed according to the manufacturer’s instructions.
Article Title: Modulatory effects of GLT-1 enhancer, MC-100093, on glutamate uptake and associated signaling pathways in female and male alcohol preferring rats exposed to ethanol
Article Snippet: .. Proteins were transferred onto nitrocellulose PVDB membranes, and then blocked with 5% milk in Tris-buffered saline Tween-20 (1x TBST) for 30 min and incubated overnight at 4°C with one of the following antibodies: rabbit anti-GLT-1 antibody (1:1000; Abcam, Ab205248 ), rabbit anti-xCT (1:1000; Abcam, Ab175186 ), rabbit anti-mTOR (1:100, Cell Signaling Technology, 7C10), rabbit anti-p-Akt (1:1000, Cell Signaling Technology, D9E), rabbit anti-Akt (1:1000, Cell Signaling Technology, C67E7), rabbit anti-NF-κB (1:1000, Huabio, ER0815), mouse anti-IkBα (1:1000, Cell Signaling Technology, L35A5), rabbit anti-GAPDH antibody (1:1000, Cell Signaling Technology, D16H11), or rabbit anti-Lamin (1:1000, Huabio, ET1606-27). .. On the following day, membranes were washed for 30 min with 1x TBST buffer and incubated with the corresponding secondary antibody (1:4000) for 1 h. Protein density was measured using ChemiDoc imaging system (Bio-Rad, USA) following incubation with chemiluminescent reagents (Super Signal West Pico, Pierce Inc.).
Article Title: MicroRNA-15a-5p suppresses triple-negative breast cancer cell proliferation and invasion by modulating T-cell immunoreceptor associated with immunoglobulin and ITIM domain expression
Article Snippet: The membrane was incubated with 5% skim milk for blocking and probed with anti-TIGIT antibodies (rabbit, 1:2000, #99567, Cell Signaling Technology, USA) overnight at 4°C. .. Following three washes with Tris-buffered saline-Tween 20, the membrane was incubated with secondary antibodies (1:10000, #58802, Cell Signaling Technology, USA) for 1.5 h under ambient conditions. .. The membrane was washed three times, and protein bands were developed using Tanon 2500 (Bio-Rad, Shanghai, China) chemiluminescence imaging system.
Article Title: Modulatory effects of GLT-1 enhancer, MC-100093, on glutamate uptake and associated signaling pathways in female and male alcohol preferring rats exposed to ethanol.
Article Snippet: .. Proteins were transferred onto nitrocellulose PVDB membranes, and then blocked with 5% milk in Tris-buffered saline Tween-20 (1x TBST) for 30 minutes and incubated overnight at 4°C with one of the following antibodies: rabbit anti-GLT-1 antibody (1:1000; Abcam, Ab205248), rabbit anti-xCT (1:1000; Abcam, Ab175186), rabbit antimTOR (1:100, Cell Signaling Technology, 7C10), rabbit anti-phospho-Akt (1:1000, Cell Signaling Technology, D9E), rabbit anti-Akt (1:1000, Cell Signaling Technology, C67E7), rabbit anti-NFkB (1:1000, Huabio, ER0815), mouse anti-IkBα (1:1000, Cell Signaling Technology, L35A5), rabbit anti-GAPDH antibody (1:1000, Cell Signaling Technology, D16H11), or rabbit anti-Lamin (1:1000, Huabio, ET1606-27). ..
Article Title: NONO as a Sensor of Intracellular Oxidation: Relevance to Neuroblastoma Cell Death
Article Snippet: .. After washing with Tris-buffered saline-Tween-20, membranes were incubated with horseradish peroxidase-conjugated secondary antibodies (1:1000, Cell Signaling Technology, Danvers, MA) for 1 h. Signals were detected with Enhanced Chemolumiscence reagent (Thermo Fisher, Waltham, MA) and captured using a gel documentation system ChemiDocTM Touch (BioRad, Hercules, CA) with multiple exposure times to ensure linear range of detection. .. The primary antibodies were: NONO (PAJ818Hu01, Cloud-Clone Corp., Houston, TX), HAND2 (E-AB-61488, Elabscience, China), MYCN, PHOX2B, β-actin (AF5204, DF9730, AF7018; Affinity Bioscience, China).
Incubation:Article Title: A preclinical and phase I clinical study of ex vivo-expanded amyloid beta-specific human regulatory T cells in Alzheimer's disease.
Article Snippet: .. Each blot was blocked for 30 min in Tris-buffered saline/Tween-20 (10 mM Tris base, 150 mM NaCl, 0.05 % Tween-20) containing 5 % fat-free milk and incubated with the following primary antibodies: CD86 (1:1000 dilution, Cell Signaling Technology), NOS2 (1:2000 dilution, Santa Cruz Biotechnology), TREM2 (1:1000 dilution, MAB1828 R&D Systems), or β-actin (1:5000 dilution, Santa Cruz Biotechnology), overnight at 4°C. ..
Article Title: H2AX phosphorylation driven by the SET-PP2A axis maintains glioma stem cell properties
Article Snippet: .. The membrane was blocked with 5% nonfat 174 milk in Tris-buffered saline-Tween 20 and incubated overnight at 4 °C with the following 175 primary antibodies: Rabbit anti-SOX2 (CST, Cat. #3579S, 1:1,000), Rabbit anti-OLIG2 176 (CST, Cat. #2748, 1:1000), Rabbit anti-NANOG (CST, Cat. #4903, 1:1,000), Mouse anti-177 GFAP (BioLegend, Cat. #644702, 1:1,000), Rabbit anti-H2AX (CST, Cat. #7631, 178 1:1,000), mouse anti-γH2AX (Millipore Sigma, Cat. #05-636, 1: 2,000), Rabbit anti-179 Tubulin (CST, Cat. #2144, 1:1,000), Rabbit anti-GAPDH (CST, Cat. #2118, 1:1,000), 180 Rabbit anti-ATM (CST, Cat. #2873, 1:1,000), Rabbit anti-pATM (S1981) (CST, Cat. 181 #5883, 1:1,000), Rabbit anti-ATR (CST, Cat. #13934, 1:1000), Rabbit anti-pATR (S428) 182 Jo rna l P repro of 9 (CST, Cat. #2853, 1:1000), Rabbit anti-pRPA32 (S33) (CST, Cat. #10148, 1:1,000), 183 Rabbit anti-RPA32 (CST, Cat. #35869, 1:1,000), Rabbit anti-SET/TAF1 antibody 184 (ABclonal, Cat. #A9173, 1:500), mouse anti-I2PP2A (SET) antibody (Santa Cruz 185 Biotechnology, Cat. #sc-133138, 1:500). .. Following multiple washes, the membrane was 186 incubated for 1 h at room temperature with HRP-conjugated goat anti-rabbit or goat anti-187 mouse secondary antibodies (Millipore Sigma, Cat. #12-348, 12-349).
Article Title: Regulation of skeletal muscle development and metabolism in broiler chickens by Urolithin A through threonine kinase 1 pathway activation
Article Snippet: .. After blocking the membrane with 5% (wt/vol) skim milk in Tris-buffered saline/Tween 20 for 1 h at 37°C, incubation was carried out overnight at 4°C with GAPDH (Proteintech, Wuhan, China), Akt1 (Cell Signaling Technology, Massachusetts, USA; ), and p-Akt1 (Cell Signaling Technology, Massachusetts, USA) antibodies. .. Subsequently, incubation with a peroxidase-conjugated anti-rabbit IgG antibody (Proteintech, Wuhan, China) was performed according to the manufacturer’s instructions.
Article Title: Modulatory effects of GLT-1 enhancer, MC-100093, on glutamate uptake and associated signaling pathways in female and male alcohol preferring rats exposed to ethanol
Article Snippet: .. Proteins were transferred onto nitrocellulose PVDB membranes, and then blocked with 5% milk in Tris-buffered saline Tween-20 (1x TBST) for 30 min and incubated overnight at 4°C with one of the following antibodies: rabbit anti-GLT-1 antibody (1:1000; Abcam, Ab205248 ), rabbit anti-xCT (1:1000; Abcam, Ab175186 ), rabbit anti-mTOR (1:100, Cell Signaling Technology, 7C10), rabbit anti-p-Akt (1:1000, Cell Signaling Technology, D9E), rabbit anti-Akt (1:1000, Cell Signaling Technology, C67E7), rabbit anti-NF-κB (1:1000, Huabio, ER0815), mouse anti-IkBα (1:1000, Cell Signaling Technology, L35A5), rabbit anti-GAPDH antibody (1:1000, Cell Signaling Technology, D16H11), or rabbit anti-Lamin (1:1000, Huabio, ET1606-27). .. On the following day, membranes were washed for 30 min with 1x TBST buffer and incubated with the corresponding secondary antibody (1:4000) for 1 h. Protein density was measured using ChemiDoc imaging system (Bio-Rad, USA) following incubation with chemiluminescent reagents (Super Signal West Pico, Pierce Inc.).
Article Title: MicroRNA-15a-5p suppresses triple-negative breast cancer cell proliferation and invasion by modulating T-cell immunoreceptor associated with immunoglobulin and ITIM domain expression
Article Snippet: The membrane was incubated with 5% skim milk for blocking and probed with anti-TIGIT antibodies (rabbit, 1:2000, #99567, Cell Signaling Technology, USA) overnight at 4°C. .. Following three washes with Tris-buffered saline-Tween 20, the membrane was incubated with secondary antibodies (1:10000, #58802, Cell Signaling Technology, USA) for 1.5 h under ambient conditions. .. The membrane was washed three times, and protein bands were developed using Tanon 2500 (Bio-Rad, Shanghai, China) chemiluminescence imaging system.
Article Title: Modulatory effects of GLT-1 enhancer, MC-100093, on glutamate uptake and associated signaling pathways in female and male alcohol preferring rats exposed to ethanol.
Article Snippet: .. Proteins were transferred onto nitrocellulose PVDB membranes, and then blocked with 5% milk in Tris-buffered saline Tween-20 (1x TBST) for 30 minutes and incubated overnight at 4°C with one of the following antibodies: rabbit anti-GLT-1 antibody (1:1000; Abcam, Ab205248), rabbit anti-xCT (1:1000; Abcam, Ab175186), rabbit antimTOR (1:100, Cell Signaling Technology, 7C10), rabbit anti-phospho-Akt (1:1000, Cell Signaling Technology, D9E), rabbit anti-Akt (1:1000, Cell Signaling Technology, C67E7), rabbit anti-NFkB (1:1000, Huabio, ER0815), mouse anti-IkBα (1:1000, Cell Signaling Technology, L35A5), rabbit anti-GAPDH antibody (1:1000, Cell Signaling Technology, D16H11), or rabbit anti-Lamin (1:1000, Huabio, ET1606-27). ..
Article Title: NONO as a Sensor of Intracellular Oxidation: Relevance to Neuroblastoma Cell Death
Article Snippet: .. After washing with Tris-buffered saline-Tween-20, membranes were incubated with horseradish peroxidase-conjugated secondary antibodies (1:1000, Cell Signaling Technology, Danvers, MA) for 1 h. Signals were detected with Enhanced Chemolumiscence reagent (Thermo Fisher, Waltham, MA) and captured using a gel documentation system ChemiDocTM Touch (BioRad, Hercules, CA) with multiple exposure times to ensure linear range of detection. .. The primary antibodies were: NONO (PAJ818Hu01, Cloud-Clone Corp., Houston, TX), HAND2 (E-AB-61488, Elabscience, China), MYCN, PHOX2B, β-actin (AF5204, DF9730, AF7018; Affinity Bioscience, China).
Membrane:Article Title: H2AX phosphorylation driven by the SET-PP2A axis maintains glioma stem cell properties
Article Snippet: .. The membrane was blocked with 5% nonfat 174 milk in Tris-buffered saline-Tween 20 and incubated overnight at 4 °C with the following 175 primary antibodies: Rabbit anti-SOX2 (CST, Cat. #3579S, 1:1,000), Rabbit anti-OLIG2 176 (CST, Cat. #2748, 1:1000), Rabbit anti-NANOG (CST, Cat. #4903, 1:1,000), Mouse anti-177 GFAP (BioLegend, Cat. #644702, 1:1,000), Rabbit anti-H2AX (CST, Cat. #7631, 178 1:1,000), mouse anti-γH2AX (Millipore Sigma, Cat. #05-636, 1: 2,000), Rabbit anti-179 Tubulin (CST, Cat. #2144, 1:1,000), Rabbit anti-GAPDH (CST, Cat. #2118, 1:1,000), 180 Rabbit anti-ATM (CST, Cat. #2873, 1:1,000), Rabbit anti-pATM (S1981) (CST, Cat. 181 #5883, 1:1,000), Rabbit anti-ATR (CST, Cat. #13934, 1:1000), Rabbit anti-pATR (S428) 182 Jo rna l P repro of 9 (CST, Cat. #2853, 1:1000), Rabbit anti-pRPA32 (S33) (CST, Cat. #10148, 1:1,000), 183 Rabbit anti-RPA32 (CST, Cat. #35869, 1:1,000), Rabbit anti-SET/TAF1 antibody 184 (ABclonal, Cat. #A9173, 1:500), mouse anti-I2PP2A (SET) antibody (Santa Cruz 185 Biotechnology, Cat. #sc-133138, 1:500). .. Following multiple washes, the membrane was 186 incubated for 1 h at room temperature with HRP-conjugated goat anti-rabbit or goat anti-187 mouse secondary antibodies (Millipore Sigma, Cat. #12-348, 12-349).
Article Title: Regulation of skeletal muscle development and metabolism in broiler chickens by Urolithin A through threonine kinase 1 pathway activation
Article Snippet: .. After blocking the membrane with 5% (wt/vol) skim milk in Tris-buffered saline/Tween 20 for 1 h at 37°C, incubation was carried out overnight at 4°C with GAPDH (Proteintech, Wuhan, China), Akt1 (Cell Signaling Technology, Massachusetts, USA; ), and p-Akt1 (Cell Signaling Technology, Massachusetts, USA) antibodies. .. Subsequently, incubation with a peroxidase-conjugated anti-rabbit IgG antibody (Proteintech, Wuhan, China) was performed according to the manufacturer’s instructions.
Article Title: MicroRNA-15a-5p suppresses triple-negative breast cancer cell proliferation and invasion by modulating T-cell immunoreceptor associated with immunoglobulin and ITIM domain expression
Article Snippet: The membrane was incubated with 5% skim milk for blocking and probed with anti-TIGIT antibodies (rabbit, 1:2000, #99567, Cell Signaling Technology, USA) overnight at 4°C. .. Following three washes with Tris-buffered saline-Tween 20, the membrane was incubated with secondary antibodies (1:10000, #58802, Cell Signaling Technology, USA) for 1.5 h under ambient conditions. .. The membrane was washed three times, and protein bands were developed using Tanon 2500 (Bio-Rad, Shanghai, China) chemiluminescence imaging system.
other:Article Title: Extracellular NCOA4 is a mediator of septic death by activating the AGER-NFKB pathway.
Article Snippet: Sepsis, a life-threatening condition resulting from a dysregulated response to pathogen infection, poses a significant challenge in clinical management.. Here, we report a novel role for the autophagy receptor NCOA4 in the pathogenesis of sepsis.. Activated macrophages and monocytes secrete NCOA4, which acts as a mediator of septic death in mice.
Blocking Assay:Article Title: Regulation of skeletal muscle development and metabolism in broiler chickens by Urolithin A through threonine kinase 1 pathway activation
Article Snippet: .. After blocking the membrane with 5% (wt/vol) skim milk in Tris-buffered saline/Tween 20 for 1 h at 37°C, incubation was carried out overnight at 4°C with GAPDH (Proteintech, Wuhan, China), Akt1 (Cell Signaling Technology, Massachusetts, USA; ), and p-Akt1 (Cell Signaling Technology, Massachusetts, USA) antibodies. .. Subsequently, incubation with a peroxidase-conjugated anti-rabbit IgG antibody (Proteintech, Wuhan, China) was performed according to the manufacturer’s instructions.
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