Chromatin Immunoprecipitation:Article Title: CRISPR-Cas3-based editing for targeted deletions in a mouse model of transthyretin amyloidosis.
Article Snippet: .. 3 nature portfolio | reporting sum m ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used All antibodies used in this study are described below with details on host species, supplier, catalog number, dilution, and validation.Immunohistochemistry (IHC):Paraffin-embedded liver sections (4 μm thick) were stained with rabbit polyclonal anti-mouse transthyretin (mTtr) antibody (1:250, LS-C407961, LSBio, Zürich, Switzerland).Frozen liver sections (4 μm thick) were stained with rabbit polyclonal anti-mTtr antibody (1:250, LS-C407961, LSBio), rabbit polyclonal anti-TTR antibody (1:1,000, 201630-T10, Sino Biological, Peking, China), and mouse monoclonal anti-CD68 antibody (1:50, ab53444, Abcam, Cambridge, UK).Western blotting (WB):Caspase-3 (Cas3) was detected with rat polyclonal anti-Cas3 antibody (1:1,000, C4U Corporation), followed by goat anti-rat IgG, HRP-linked antibody (1:2,000, #7077, Cell Signaling Technology, Danvers, MA, USA).α-Tubulin was detected using rabbit monoclonal α-Tubulin antibody (1:2,000, #2144, Cell Signaling Technology), followed by goat anti-rabbit IgG, HRP-linked antibody (1:2,000, #7074, Cell Signaling Technology). .. Validation Validation of all primary antibodies was confirmed by the manufacturers and supported by experimental results described in the Methods section (“Immunohistochemistry” and “Western blotting”).The anti-mTtr (LS-C407961, LSBio) and anti-TTR (201630-T10, Sino Biological) antibodies produced specific cytoplasmic staining in hepatocytes consistent with the known localization of transthyretin, in agreement with previous reports and manufacturer datasheets.The anti-CD68 antibody (ab53444, Abcam) selectively labeled macrophages in hepatic tissue, consistent with the manufacturer’s validation data.The anti-Cas3 antibody (C4U Corporation) yielded a single band at the expected molecular weight in Western blotting, and recombinant Cas3 protein was used as a positive control to confirm antibody specificity.The α-Tubulin antibody (#2144, Cell Signaling) produced a single band corresponding to the expected molecular weight, as validated by the manufacturer.All antibodies showed staining or banding patterns consistent with the expected expression profiles in mouse liver samples.
Flow Cytometry:Article Title: CRISPR-Cas3-based editing for targeted deletions in a mouse model of transthyretin amyloidosis.
Article Snippet: .. 3 nature portfolio | reporting sum m ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used All antibodies used in this study are described below with details on host species, supplier, catalog number, dilution, and validation.Immunohistochemistry (IHC):Paraffin-embedded liver sections (4 μm thick) were stained with rabbit polyclonal anti-mouse transthyretin (mTtr) antibody (1:250, LS-C407961, LSBio, Zürich, Switzerland).Frozen liver sections (4 μm thick) were stained with rabbit polyclonal anti-mTtr antibody (1:250, LS-C407961, LSBio), rabbit polyclonal anti-TTR antibody (1:1,000, 201630-T10, Sino Biological, Peking, China), and mouse monoclonal anti-CD68 antibody (1:50, ab53444, Abcam, Cambridge, UK).Western blotting (WB):Caspase-3 (Cas3) was detected with rat polyclonal anti-Cas3 antibody (1:1,000, C4U Corporation), followed by goat anti-rat IgG, HRP-linked antibody (1:2,000, #7077, Cell Signaling Technology, Danvers, MA, USA).α-Tubulin was detected using rabbit monoclonal α-Tubulin antibody (1:2,000, #2144, Cell Signaling Technology), followed by goat anti-rabbit IgG, HRP-linked antibody (1:2,000, #7074, Cell Signaling Technology). .. Validation Validation of all primary antibodies was confirmed by the manufacturers and supported by experimental results described in the Methods section (“Immunohistochemistry” and “Western blotting”).The anti-mTtr (LS-C407961, LSBio) and anti-TTR (201630-T10, Sino Biological) antibodies produced specific cytoplasmic staining in hepatocytes consistent with the known localization of transthyretin, in agreement with previous reports and manufacturer datasheets.The anti-CD68 antibody (ab53444, Abcam) selectively labeled macrophages in hepatic tissue, consistent with the manufacturer’s validation data.The anti-Cas3 antibody (C4U Corporation) yielded a single band at the expected molecular weight in Western blotting, and recombinant Cas3 protein was used as a positive control to confirm antibody specificity.The α-Tubulin antibody (#2144, Cell Signaling) produced a single band corresponding to the expected molecular weight, as validated by the manufacturer.All antibodies showed staining or banding patterns consistent with the expected expression profiles in mouse liver samples.
Magnetic Resonance Imaging:Article Title: CRISPR-Cas3-based editing for targeted deletions in a mouse model of transthyretin amyloidosis.
Article Snippet: .. 3 nature portfolio | reporting sum m ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used All antibodies used in this study are described below with details on host species, supplier, catalog number, dilution, and validation.Immunohistochemistry (IHC):Paraffin-embedded liver sections (4 μm thick) were stained with rabbit polyclonal anti-mouse transthyretin (mTtr) antibody (1:250, LS-C407961, LSBio, Zürich, Switzerland).Frozen liver sections (4 μm thick) were stained with rabbit polyclonal anti-mTtr antibody (1:250, LS-C407961, LSBio), rabbit polyclonal anti-TTR antibody (1:1,000, 201630-T10, Sino Biological, Peking, China), and mouse monoclonal anti-CD68 antibody (1:50, ab53444, Abcam, Cambridge, UK).Western blotting (WB):Caspase-3 (Cas3) was detected with rat polyclonal anti-Cas3 antibody (1:1,000, C4U Corporation), followed by goat anti-rat IgG, HRP-linked antibody (1:2,000, #7077, Cell Signaling Technology, Danvers, MA, USA).α-Tubulin was detected using rabbit monoclonal α-Tubulin antibody (1:2,000, #2144, Cell Signaling Technology), followed by goat anti-rabbit IgG, HRP-linked antibody (1:2,000, #7074, Cell Signaling Technology). .. Validation Validation of all primary antibodies was confirmed by the manufacturers and supported by experimental results described in the Methods section (“Immunohistochemistry” and “Western blotting”).The anti-mTtr (LS-C407961, LSBio) and anti-TTR (201630-T10, Sino Biological) antibodies produced specific cytoplasmic staining in hepatocytes consistent with the known localization of transthyretin, in agreement with previous reports and manufacturer datasheets.The anti-CD68 antibody (ab53444, Abcam) selectively labeled macrophages in hepatic tissue, consistent with the manufacturer’s validation data.The anti-Cas3 antibody (C4U Corporation) yielded a single band at the expected molecular weight in Western blotting, and recombinant Cas3 protein was used as a positive control to confirm antibody specificity.The α-Tubulin antibody (#2144, Cell Signaling) produced a single band corresponding to the expected molecular weight, as validated by the manufacturer.All antibodies showed staining or banding patterns consistent with the expected expression profiles in mouse liver samples.
Biomarker Discovery:Article Title: CRISPR-Cas3-based editing for targeted deletions in a mouse model of transthyretin amyloidosis.
Article Snippet: .. 3 nature portfolio | reporting sum m ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used All antibodies used in this study are described below with details on host species, supplier, catalog number, dilution, and validation.Immunohistochemistry (IHC):Paraffin-embedded liver sections (4 μm thick) were stained with rabbit polyclonal anti-mouse transthyretin (mTtr) antibody (1:250, LS-C407961, LSBio, Zürich, Switzerland).Frozen liver sections (4 μm thick) were stained with rabbit polyclonal anti-mTtr antibody (1:250, LS-C407961, LSBio), rabbit polyclonal anti-TTR antibody (1:1,000, 201630-T10, Sino Biological, Peking, China), and mouse monoclonal anti-CD68 antibody (1:50, ab53444, Abcam, Cambridge, UK).Western blotting (WB):Caspase-3 (Cas3) was detected with rat polyclonal anti-Cas3 antibody (1:1,000, C4U Corporation), followed by goat anti-rat IgG, HRP-linked antibody (1:2,000, #7077, Cell Signaling Technology, Danvers, MA, USA).α-Tubulin was detected using rabbit monoclonal α-Tubulin antibody (1:2,000, #2144, Cell Signaling Technology), followed by goat anti-rabbit IgG, HRP-linked antibody (1:2,000, #7074, Cell Signaling Technology). .. Validation Validation of all primary antibodies was confirmed by the manufacturers and supported by experimental results described in the Methods section (“Immunohistochemistry” and “Western blotting”).The anti-mTtr (LS-C407961, LSBio) and anti-TTR (201630-T10, Sino Biological) antibodies produced specific cytoplasmic staining in hepatocytes consistent with the known localization of transthyretin, in agreement with previous reports and manufacturer datasheets.The anti-CD68 antibody (ab53444, Abcam) selectively labeled macrophages in hepatic tissue, consistent with the manufacturer’s validation data.The anti-Cas3 antibody (C4U Corporation) yielded a single band at the expected molecular weight in Western blotting, and recombinant Cas3 protein was used as a positive control to confirm antibody specificity.The α-Tubulin antibody (#2144, Cell Signaling) produced a single band corresponding to the expected molecular weight, as validated by the manufacturer.All antibodies showed staining or banding patterns consistent with the expected expression profiles in mouse liver samples.
Immunohistochemistry:Article Title: CRISPR-Cas3-based editing for targeted deletions in a mouse model of transthyretin amyloidosis.
Article Snippet: .. 3 nature portfolio | reporting sum m ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used All antibodies used in this study are described below with details on host species, supplier, catalog number, dilution, and validation.Immunohistochemistry (IHC):Paraffin-embedded liver sections (4 μm thick) were stained with rabbit polyclonal anti-mouse transthyretin (mTtr) antibody (1:250, LS-C407961, LSBio, Zürich, Switzerland).Frozen liver sections (4 μm thick) were stained with rabbit polyclonal anti-mTtr antibody (1:250, LS-C407961, LSBio), rabbit polyclonal anti-TTR antibody (1:1,000, 201630-T10, Sino Biological, Peking, China), and mouse monoclonal anti-CD68 antibody (1:50, ab53444, Abcam, Cambridge, UK).Western blotting (WB):Caspase-3 (Cas3) was detected with rat polyclonal anti-Cas3 antibody (1:1,000, C4U Corporation), followed by goat anti-rat IgG, HRP-linked antibody (1:2,000, #7077, Cell Signaling Technology, Danvers, MA, USA).α-Tubulin was detected using rabbit monoclonal α-Tubulin antibody (1:2,000, #2144, Cell Signaling Technology), followed by goat anti-rabbit IgG, HRP-linked antibody (1:2,000, #7074, Cell Signaling Technology). .. Validation Validation of all primary antibodies was confirmed by the manufacturers and supported by experimental results described in the Methods section (“Immunohistochemistry” and “Western blotting”).The anti-mTtr (LS-C407961, LSBio) and anti-TTR (201630-T10, Sino Biological) antibodies produced specific cytoplasmic staining in hepatocytes consistent with the known localization of transthyretin, in agreement with previous reports and manufacturer datasheets.The anti-CD68 antibody (ab53444, Abcam) selectively labeled macrophages in hepatic tissue, consistent with the manufacturer’s validation data.The anti-Cas3 antibody (C4U Corporation) yielded a single band at the expected molecular weight in Western blotting, and recombinant Cas3 protein was used as a positive control to confirm antibody specificity.The α-Tubulin antibody (#2144, Cell Signaling) produced a single band corresponding to the expected molecular weight, as validated by the manufacturer.All antibodies showed staining or banding patterns consistent with the expected expression profiles in mouse liver samples.
Staining:Article Title: CRISPR-Cas3-based editing for targeted deletions in a mouse model of transthyretin amyloidosis.
Article Snippet: .. 3 nature portfolio | reporting sum m ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used All antibodies used in this study are described below with details on host species, supplier, catalog number, dilution, and validation.Immunohistochemistry (IHC):Paraffin-embedded liver sections (4 μm thick) were stained with rabbit polyclonal anti-mouse transthyretin (mTtr) antibody (1:250, LS-C407961, LSBio, Zürich, Switzerland).Frozen liver sections (4 μm thick) were stained with rabbit polyclonal anti-mTtr antibody (1:250, LS-C407961, LSBio), rabbit polyclonal anti-TTR antibody (1:1,000, 201630-T10, Sino Biological, Peking, China), and mouse monoclonal anti-CD68 antibody (1:50, ab53444, Abcam, Cambridge, UK).Western blotting (WB):Caspase-3 (Cas3) was detected with rat polyclonal anti-Cas3 antibody (1:1,000, C4U Corporation), followed by goat anti-rat IgG, HRP-linked antibody (1:2,000, #7077, Cell Signaling Technology, Danvers, MA, USA).α-Tubulin was detected using rabbit monoclonal α-Tubulin antibody (1:2,000, #2144, Cell Signaling Technology), followed by goat anti-rabbit IgG, HRP-linked antibody (1:2,000, #7074, Cell Signaling Technology). .. Validation Validation of all primary antibodies was confirmed by the manufacturers and supported by experimental results described in the Methods section (“Immunohistochemistry” and “Western blotting”).The anti-mTtr (LS-C407961, LSBio) and anti-TTR (201630-T10, Sino Biological) antibodies produced specific cytoplasmic staining in hepatocytes consistent with the known localization of transthyretin, in agreement with previous reports and manufacturer datasheets.The anti-CD68 antibody (ab53444, Abcam) selectively labeled macrophages in hepatic tissue, consistent with the manufacturer’s validation data.The anti-Cas3 antibody (C4U Corporation) yielded a single band at the expected molecular weight in Western blotting, and recombinant Cas3 protein was used as a positive control to confirm antibody specificity.The α-Tubulin antibody (#2144, Cell Signaling) produced a single band corresponding to the expected molecular weight, as validated by the manufacturer.All antibodies showed staining or banding patterns consistent with the expected expression profiles in mouse liver samples.
Western Blot:Article Title: CRISPR-Cas3-based editing for targeted deletions in a mouse model of transthyretin amyloidosis.
Article Snippet: .. 3 nature portfolio | reporting sum m ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used All antibodies used in this study are described below with details on host species, supplier, catalog number, dilution, and validation.Immunohistochemistry (IHC):Paraffin-embedded liver sections (4 μm thick) were stained with rabbit polyclonal anti-mouse transthyretin (mTtr) antibody (1:250, LS-C407961, LSBio, Zürich, Switzerland).Frozen liver sections (4 μm thick) were stained with rabbit polyclonal anti-mTtr antibody (1:250, LS-C407961, LSBio), rabbit polyclonal anti-TTR antibody (1:1,000, 201630-T10, Sino Biological, Peking, China), and mouse monoclonal anti-CD68 antibody (1:50, ab53444, Abcam, Cambridge, UK).Western blotting (WB):Caspase-3 (Cas3) was detected with rat polyclonal anti-Cas3 antibody (1:1,000, C4U Corporation), followed by goat anti-rat IgG, HRP-linked antibody (1:2,000, #7077, Cell Signaling Technology, Danvers, MA, USA).α-Tubulin was detected using rabbit monoclonal α-Tubulin antibody (1:2,000, #2144, Cell Signaling Technology), followed by goat anti-rabbit IgG, HRP-linked antibody (1:2,000, #7074, Cell Signaling Technology). .. Validation Validation of all primary antibodies was confirmed by the manufacturers and supported by experimental results described in the Methods section (“Immunohistochemistry” and “Western blotting”).The anti-mTtr (LS-C407961, LSBio) and anti-TTR (201630-T10, Sino Biological) antibodies produced specific cytoplasmic staining in hepatocytes consistent with the known localization of transthyretin, in agreement with previous reports and manufacturer datasheets.The anti-CD68 antibody (ab53444, Abcam) selectively labeled macrophages in hepatic tissue, consistent with the manufacturer’s validation data.The anti-Cas3 antibody (C4U Corporation) yielded a single band at the expected molecular weight in Western blotting, and recombinant Cas3 protein was used as a positive control to confirm antibody specificity.The α-Tubulin antibody (#2144, Cell Signaling) produced a single band corresponding to the expected molecular weight, as validated by the manufacturer.All antibodies showed staining or banding patterns consistent with the expected expression profiles in mouse liver samples.
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