Staining:Article Title: RANK drives structured intestinal epithelial expansion during pregnancy
Article Snippet: Confocal images were obtained using the Zeiss LSM 700 and Zeiss LSM 710 microscopes. .. For the whole-mount staining of mouse and human organoids, organoids were fixed with 4% PFA at room temperature for 15 min, followed by incubation with blocking and permeabilization solution consisting of 0.2% Triton X-100, 0.1% Tween-20, 2% BSA and 2% normal goat serum in PBS at room temperature for 1 h. Mouse organoids were stained at 4 °C overnight with anti-mouse OLFM4 (1:400; Cell Signaling Technology, 39141) in blocking and permeabilization solution. .. Goat anti-rabbit Alexa Fluor 633 (1:500; Invitrogen, A21072) was used as a secondary antibody.
Article Title: RANK drives structured intestinal epithelial expansion during pregnancy.
Article Snippet: Confocal images were obtained using the Zeiss LSM 700 and Zeiss LSM 710 microscopes. .. For the whole-mount staining of mouse and human organoids, organoids were fixed with 4% PFA at room temperature for 15 min, followed by incubation with blocking and permeabilization solution consisting of 0.2% Triton X-100, 0.1% Tween-20, 2% BSA and 2% normal goat serum in PBS at room temperature for 1 h. Mouse organoids were stained at 4 °C overnight with anti-mouse OLFM4 (1:400; Cell Signaling Technology, 39141) in blocking and permeabilization solution. .. After dead cells were fluorescently labelled with a viability dye (eBioscience, 1:1,200), dissociated cells were fixed, permeabilized using a fixation/permeabilization kit (eBioscience) and finally stained using the Click-iT EdU kit (Life Technologies).
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 Propidium Iodide Flow Cytometry Kit (ab139418, Abcam) was used to distinguish cell cycle stages on basis of the cellular DNA content, following the manufacturer's protocol.
Incubation:Article Title: RANK drives structured intestinal epithelial expansion during pregnancy
Article Snippet: Confocal images were obtained using the Zeiss LSM 700 and Zeiss LSM 710 microscopes. .. For the whole-mount staining of mouse and human organoids, organoids were fixed with 4% PFA at room temperature for 15 min, followed by incubation with blocking and permeabilization solution consisting of 0.2% Triton X-100, 0.1% Tween-20, 2% BSA and 2% normal goat serum in PBS at room temperature for 1 h. Mouse organoids were stained at 4 °C overnight with anti-mouse OLFM4 (1:400; Cell Signaling Technology, 39141) in blocking and permeabilization solution. .. Goat anti-rabbit Alexa Fluor 633 (1:500; Invitrogen, A21072) was used as a secondary antibody.
Article Title: RANK drives structured intestinal epithelial expansion during pregnancy.
Article Snippet: Confocal images were obtained using the Zeiss LSM 700 and Zeiss LSM 710 microscopes. .. For the whole-mount staining of mouse and human organoids, organoids were fixed with 4% PFA at room temperature for 15 min, followed by incubation with blocking and permeabilization solution consisting of 0.2% Triton X-100, 0.1% Tween-20, 2% BSA and 2% normal goat serum in PBS at room temperature for 1 h. Mouse organoids were stained at 4 °C overnight with anti-mouse OLFM4 (1:400; Cell Signaling Technology, 39141) in blocking and permeabilization solution. .. After dead cells were fluorescently labelled with a viability dye (eBioscience, 1:1,200), dissociated cells were fixed, permeabilized using a fixation/permeabilization kit (eBioscience) and finally stained using the Click-iT EdU kit (Life Technologies).
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 conserved cysteine in the DNA-binding domain of MmuPV1 E2 is required for replication in vivo
Article Snippet: .. Permeabilization solution was removed and cells were incubated in blocking solution (PBS + 0.1% Triton-X100 + 10% normal goat serum) for approximately 1 h. Primary rabbit anti-V5 antibody (CST D3H8Q) was added at an approximate 1:1,200 dilution and incubated rocking overnight at 4°C. .. Primary antibody was removed; cells were washed 3× in PBS; and then cells were incubated with AlexaFluor 555 Goat anti-rabbit (#A21428) at a dilution of 1:1,500 for approximately 2 h at room temperature and protected from light.
Article Title: The TDG protein environment connects active DNA demethylation with chromatin and RNA biology.
Article Snippet: .. 1x106 cells were fixed with 100 μL 4% formaldehyde diluted in 323 PBS at RT for 10 min and treated with 100 μL permeabilization solution (1% BSA, 0.1% Triton 324 X-100, in PBS) at RT for 30 min. After centrifugation, cells were incubated with 100 μL HA 325 antibody (Cell Signaling C29F4, 1:800) diluted in blocking solution (PBS, 1% BSA) at RT for 326 1 h. After three washing steps, cells were incubated with the secondary Alexa-488 anti-mouse 327 (Invitrogen, 1:2’000) antibody diluted in blocking solution at RT for 30 min. .. Cells were washed 328 three times, stained with DAPI (200 ng/mL in blocking solution) at RT for 10 min, washed again 329 and analyzed with CytoFLEX (Beckman Coulter) cytometer.
Article Title: A conserved cysteine in the DNA-binding domain of MmuPV1 E2 is required for replication in vivo .
Article Snippet: The papillomavirus (PV) E2 protein is highly conserved, consisting of an N-terminal transactivation domain linked to a C-terminal DNA binding and dimerization domain (DBD) by a flexible hinge region.. The E2 DBD exhibits a helix-turn-helix structure that dimerizes into a beta barrel prior to binding DNA; the first helix, α1, is responsible for recognition of the palindromic E2 binding site.. The DNA recognition helix consists of a tract of basic amino acids with a highly conserved central cysteine residue.
Blocking Assay:Article Title: RANK drives structured intestinal epithelial expansion during pregnancy
Article Snippet: Confocal images were obtained using the Zeiss LSM 700 and Zeiss LSM 710 microscopes. .. For the whole-mount staining of mouse and human organoids, organoids were fixed with 4% PFA at room temperature for 15 min, followed by incubation with blocking and permeabilization solution consisting of 0.2% Triton X-100, 0.1% Tween-20, 2% BSA and 2% normal goat serum in PBS at room temperature for 1 h. Mouse organoids were stained at 4 °C overnight with anti-mouse OLFM4 (1:400; Cell Signaling Technology, 39141) in blocking and permeabilization solution. .. Goat anti-rabbit Alexa Fluor 633 (1:500; Invitrogen, A21072) was used as a secondary antibody.
Article Title: RANK drives structured intestinal epithelial expansion during pregnancy.
Article Snippet: Confocal images were obtained using the Zeiss LSM 700 and Zeiss LSM 710 microscopes. .. For the whole-mount staining of mouse and human organoids, organoids were fixed with 4% PFA at room temperature for 15 min, followed by incubation with blocking and permeabilization solution consisting of 0.2% Triton X-100, 0.1% Tween-20, 2% BSA and 2% normal goat serum in PBS at room temperature for 1 h. Mouse organoids were stained at 4 °C overnight with anti-mouse OLFM4 (1:400; Cell Signaling Technology, 39141) in blocking and permeabilization solution. .. After dead cells were fluorescently labelled with a viability dye (eBioscience, 1:1,200), dissociated cells were fixed, permeabilized using a fixation/permeabilization kit (eBioscience) and finally stained using the Click-iT EdU kit (Life Technologies).
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 conserved cysteine in the DNA-binding domain of MmuPV1 E2 is required for replication in vivo
Article Snippet: .. Permeabilization solution was removed and cells were incubated in blocking solution (PBS + 0.1% Triton-X100 + 10% normal goat serum) for approximately 1 h. Primary rabbit anti-V5 antibody (CST D3H8Q) was added at an approximate 1:1,200 dilution and incubated rocking overnight at 4°C. .. Primary antibody was removed; cells were washed 3× in PBS; and then cells were incubated with AlexaFluor 555 Goat anti-rabbit (#A21428) at a dilution of 1:1,500 for approximately 2 h at room temperature and protected from light.
Article Title: The TDG protein environment connects active DNA demethylation with chromatin and RNA biology.
Article Snippet: .. 1x106 cells were fixed with 100 μL 4% formaldehyde diluted in 323 PBS at RT for 10 min and treated with 100 μL permeabilization solution (1% BSA, 0.1% Triton 324 X-100, in PBS) at RT for 30 min. After centrifugation, cells were incubated with 100 μL HA 325 antibody (Cell Signaling C29F4, 1:800) diluted in blocking solution (PBS, 1% BSA) at RT for 326 1 h. After three washing steps, cells were incubated with the secondary Alexa-488 anti-mouse 327 (Invitrogen, 1:2’000) antibody diluted in blocking solution at RT for 30 min. .. Cells were washed 328 three times, stained with DAPI (200 ng/mL in blocking solution) at RT for 10 min, washed again 329 and analyzed with CytoFLEX (Beckman Coulter) cytometer.
Article Title: A conserved cysteine in the DNA-binding domain of MmuPV1 E2 is required for replication in vivo .
Article Snippet: The papillomavirus (PV) E2 protein is highly conserved, consisting of an N-terminal transactivation domain linked to a C-terminal DNA binding and dimerization domain (DBD) by a flexible hinge region.. The E2 DBD exhibits a helix-turn-helix structure that dimerizes into a beta barrel prior to binding DNA; the first helix, α1, is responsible for recognition of the palindromic E2 binding site.. The DNA recognition helix consists of a tract of basic amino acids with a highly conserved central cysteine residue.
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.
Article Title: The TDG protein environment connects active DNA demethylation with chromatin and RNA biology.
Article Snippet: .. 1x106 cells were fixed with 100 μL 4% formaldehyde diluted in 323 PBS at RT for 10 min and treated with 100 μL permeabilization solution (1% BSA, 0.1% Triton 324 X-100, in PBS) at RT for 30 min. After centrifugation, cells were incubated with 100 μL HA 325 antibody (Cell Signaling C29F4, 1:800) diluted in blocking solution (PBS, 1% BSA) at RT for 326 1 h. After three washing steps, cells were incubated with the secondary Alexa-488 anti-mouse 327 (Invitrogen, 1:2’000) antibody diluted in blocking solution at RT for 30 min. .. Cells were washed 328 three times, stained with DAPI (200 ng/mL in blocking solution) at RT for 10 min, washed again 329 and analyzed with CytoFLEX (Beckman Coulter) cytometer.
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