Incubation:Article Title: Spatial Targeting of Sialic Acid Receptors for MRI/BNCT-Integrated Boron Drug-Based Antitumor Therapy.
Article Snippet: Objective Boron neutron capture therapy (BNCT) is limited by inadequate boron delivery to tumor cells and the inability to visualize boron agents for personalized treatment.. This study introduces DOTA-BPA-Gd, a dual-function boron agent integrating BNCT efficacy with drug tracing.. Methods DOTA-BPA-Gd was designed with four phenylboronic acid groups to target sialic acid residues on cancer cell surfaces.
Article Title: The TDG protein environment connects active DNA demethylation with chromatin and RNA biology
Article Snippet: .. 1 × 10 6 cells were fixed with 100 μL 4% formaldehyde diluted in PBS at RT for 10 min and treated with 100 μL permeabilization solution (1% BSA, 0.1% Triton X-100, in PBS) at RT for 30 min. After centrifugation, cells were incubated with 100 μL HA antibody (Cell Signaling C29F4, 1:800) diluted in blocking solution (PBS, 1% BSA) at RT for 1 h. After three washing steps, cells were incubated with the secondary Alexa-488 anti-mouse (Invitrogen, 1:2′000) antibody diluted in blocking solution at RT for 30 min. .. Cells were washed three times, stained with DAPI (200 ng/mL in blocking solution) at RT for 10 min, washed again and analyzed with CytoFLEX (Beckman Coulter) cytometer.
Article Title: A novel iPSC model of Bryant-Li-Bhoj neurodevelopmental/neurodegenerative syndrome demonstrates the role of histone H3.3 in chromatin dynamics, neuronal differentiation, and maturation.
Article Snippet: .. For the flow cytometry analysis, cell permeabilization was performed with saponin buffer (diluted to 1 × in water; Biolegend, 421,022), followed by a 1hour incubation with antibodies diluted in saponin buffer: Forse-1 (DSHB, 1:100), NCAD (Cell Signaling #14215, 1:300), and an IgM isotype control (Santa Cruz, sc-3881, 1:100). .. The cells were then washed with either saponin buffer or FACS buffer and incubated for 1 h with secondary antibodies: goat anti-rabbit Alexa 488 (Jackson ImmunoResearch, 111-545-144, 1:500) or goat anti-mouse IgM-conjugated Alexa 488 (Thermo Fisher Scientific, A21042, 1:500).
Article Title: Avian GCGR-mediated continuous fat utilization offers perspectives for obesity treatment
Article Snippet: .. Slides were washed in PBS (pH = 7.4) on a decolorizing shaker for 3 times 5 min. After addition of 50–100 μl permeabilization working solution, the following steps were: incubation at room temperature for 20 min, 3 washes with PBS, each for 5 min. A few drops of blocking buffer (10% donkey serum blocking buffer for incubation with goat-derived primary antibodies, or 3% BSA blocking buffer for incubation with primary antibodies from other species) were added, and incubation was carried out at room temperature for 30 min. Pre-prepared primary antibody anti-v5 (1:200, CST, USA) was added, and the slides were incubated flat in a humidified chamber at 4 °C overnight. ..
Article Title: A novel iPSC model of Bryant-Li-Bhoj neurodevelopmental/neurodegenerative syndrome demonstrates the role of histone H3.3 in chromatin dynamics, neuronal differentiation, and maturation
Article Snippet: .. For the flow cytometry analysis, cell permeabilization was performed with saponin buffer (diluted to 1 × in water; Biolegend, 421,022), followed by a 1hour incubation with antibodies diluted in saponin buffer: Forse-1 (DSHB, 1:100), NCAD (Cell Signaling #14215, 1:300), and an IgM isotype control (Santa Cruz, sc-3881, 1:100). .. The cells were then washed with either saponin buffer or FACS buffer and incubated for 1 h with secondary antibodies: goat anti-rabbit Alexa 488 (Jackson ImmunoResearch, 111-545-144, 1:500) or goat anti-mouse IgM-conjugated Alexa 488 (Thermo Fisher Scientific, A21042, 1:500).
Centrifugation:Article Title: The TDG protein environment connects active DNA demethylation with chromatin and RNA biology
Article Snippet: .. 1 × 10 6 cells were fixed with 100 μL 4% formaldehyde diluted in PBS at RT for 10 min and treated with 100 μL permeabilization solution (1% BSA, 0.1% Triton X-100, in PBS) at RT for 30 min. After centrifugation, cells were incubated with 100 μL HA antibody (Cell Signaling C29F4, 1:800) diluted in blocking solution (PBS, 1% BSA) at RT for 1 h. After three washing steps, cells were incubated with the secondary Alexa-488 anti-mouse (Invitrogen, 1:2′000) antibody diluted in blocking solution at RT for 30 min. .. Cells were washed three times, stained with DAPI (200 ng/mL in blocking solution) at RT for 10 min, washed again and analyzed with CytoFLEX (Beckman Coulter) cytometer.
Blocking Assay:Article Title: The TDG protein environment connects active DNA demethylation with chromatin and RNA biology
Article Snippet: .. 1 × 10 6 cells were fixed with 100 μL 4% formaldehyde diluted in PBS at RT for 10 min and treated with 100 μL permeabilization solution (1% BSA, 0.1% Triton X-100, in PBS) at RT for 30 min. After centrifugation, cells were incubated with 100 μL HA antibody (Cell Signaling C29F4, 1:800) diluted in blocking solution (PBS, 1% BSA) at RT for 1 h. After three washing steps, cells were incubated with the secondary Alexa-488 anti-mouse (Invitrogen, 1:2′000) antibody diluted in blocking solution at RT for 30 min. .. Cells were washed three times, stained with DAPI (200 ng/mL in blocking solution) at RT for 10 min, washed again and analyzed with CytoFLEX (Beckman Coulter) cytometer.
Article Title: Avian GCGR-mediated continuous fat utilization offers perspectives for obesity treatment
Article Snippet: .. Slides were washed in PBS (pH = 7.4) on a decolorizing shaker for 3 times 5 min. After addition of 50–100 μl permeabilization working solution, the following steps were: incubation at room temperature for 20 min, 3 washes with PBS, each for 5 min. A few drops of blocking buffer (10% donkey serum blocking buffer for incubation with goat-derived primary antibodies, or 3% BSA blocking buffer for incubation with primary antibodies from other species) were added, and incubation was carried out at room temperature for 30 min. Pre-prepared primary antibody anti-v5 (1:200, CST, USA) was added, and the slides were incubated flat in a humidified chamber at 4 °C overnight. ..
Flow Cytometry:Article Title: A novel iPSC model of Bryant-Li-Bhoj neurodevelopmental/neurodegenerative syndrome demonstrates the role of histone H3.3 in chromatin dynamics, neuronal differentiation, and maturation.
Article Snippet: .. For the flow cytometry analysis, cell permeabilization was performed with saponin buffer (diluted to 1 × in water; Biolegend, 421,022), followed by a 1hour incubation with antibodies diluted in saponin buffer: Forse-1 (DSHB, 1:100), NCAD (Cell Signaling #14215, 1:300), and an IgM isotype control (Santa Cruz, sc-3881, 1:100). .. The cells were then washed with either saponin buffer or FACS buffer and incubated for 1 h with secondary antibodies: goat anti-rabbit Alexa 488 (Jackson ImmunoResearch, 111-545-144, 1:500) or goat anti-mouse IgM-conjugated Alexa 488 (Thermo Fisher Scientific, A21042, 1:500).
Article Title: A novel iPSC model of Bryant-Li-Bhoj neurodevelopmental/neurodegenerative syndrome demonstrates the role of histone H3.3 in chromatin dynamics, neuronal differentiation, and maturation
Article Snippet: .. For the flow cytometry analysis, cell permeabilization was performed with saponin buffer (diluted to 1 × in water; Biolegend, 421,022), followed by a 1hour incubation with antibodies diluted in saponin buffer: Forse-1 (DSHB, 1:100), NCAD (Cell Signaling #14215, 1:300), and an IgM isotype control (Santa Cruz, sc-3881, 1:100). .. The cells were then washed with either saponin buffer or FACS buffer and incubated for 1 h with secondary antibodies: goat anti-rabbit Alexa 488 (Jackson ImmunoResearch, 111-545-144, 1:500) or goat anti-mouse IgM-conjugated Alexa 488 (Thermo Fisher Scientific, A21042, 1:500).
Control:Article Title: A novel iPSC model of Bryant-Li-Bhoj neurodevelopmental/neurodegenerative syndrome demonstrates the role of histone H3.3 in chromatin dynamics, neuronal differentiation, and maturation.
Article Snippet: .. For the flow cytometry analysis, cell permeabilization was performed with saponin buffer (diluted to 1 × in water; Biolegend, 421,022), followed by a 1hour incubation with antibodies diluted in saponin buffer: Forse-1 (DSHB, 1:100), NCAD (Cell Signaling #14215, 1:300), and an IgM isotype control (Santa Cruz, sc-3881, 1:100). .. The cells were then washed with either saponin buffer or FACS buffer and incubated for 1 h with secondary antibodies: goat anti-rabbit Alexa 488 (Jackson ImmunoResearch, 111-545-144, 1:500) or goat anti-mouse IgM-conjugated Alexa 488 (Thermo Fisher Scientific, A21042, 1:500).
Article Title: A novel iPSC model of Bryant-Li-Bhoj neurodevelopmental/neurodegenerative syndrome demonstrates the role of histone H3.3 in chromatin dynamics, neuronal differentiation, and maturation
Article Snippet: .. For the flow cytometry analysis, cell permeabilization was performed with saponin buffer (diluted to 1 × in water; Biolegend, 421,022), followed by a 1hour incubation with antibodies diluted in saponin buffer: Forse-1 (DSHB, 1:100), NCAD (Cell Signaling #14215, 1:300), and an IgM isotype control (Santa Cruz, sc-3881, 1:100). .. The cells were then washed with either saponin buffer or FACS buffer and incubated for 1 h with secondary antibodies: goat anti-rabbit Alexa 488 (Jackson ImmunoResearch, 111-545-144, 1:500) or goat anti-mouse IgM-conjugated Alexa 488 (Thermo Fisher Scientific, A21042, 1:500).
Staining:Article Title: The type-I interferon priming signal balances antibacterial and antitumor trained immunity in alveolar macrophages.
Article Snippet: Tissue-resident macrophages may be trained to confer an enhanced response to heterologous restimulation and thus foster versatile trained immunity (TI) against both infections and tumors.. However, the key priming signals that contribute to such functional versatility in trained macrophages are not fully understood.. Here, we show that influenza A virus (IAV) infection in mice induces lasting transcriptional imprints of acute type-I interferon (IFN-I) signaling in lung-resident alveolar macrophages (AMs) that confer balanced antibacterial and antitumor TI responses.
Article Title: Boron neutron capture therapy preserves immune cells and induces robust anti-tumour immunity in preclinical mouse model.
Article Snippet: .. The sections were fixed with 4% paraformaldehyde and treated with a permeabilization solution containing 0.5% Triton X-100 for 30 minutes at room temperature, and then stained with the following primary antibodies: phospho-histone H2A.X (Ser139) antibody (9718S, CST, 1:200, clone number 20E3), anti-HMGB1 polyclonal antibody (ab18256, Abcam; 1:200), anti-Calreticulin polyclonal antibody (ab2907, Abcam, 1:200) or CD8a monoclonal antibody (14- 0081-82, Invitrogen; 1:200, clone number 53-6.7), as well as the secondary antibody anti-rabbit IgG (H+L) (4412S, CST; 1:1000). .. The samples were mounted with anti-fluorescence quenching sealing tablets (G1407, Servicebio) containing 4',6-diamidino-2-phenylindole (DAPI) and then imaged using a fluorescence microscope (IX73, Olympus).
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