Western Blot:Article Title: Pre-established ATF4 occupancy and chromatin organization instruct selective transcription activation during integrated stress response
Article Snippet: .. This is a study with data collection and analyses relying on objective measures. anti-rabbit ATF4 (Cell Signaling Technology #11815; 1:1000 dilution for Western Blot; 1:200 dilution for immunouorescence; 1:50 dilution for CUT&Tag; 1:50 dilution for immunoprecipitation ) anti-mouse ATF4 (Santa Cruz Biotechnology #sc-390063,1:50 dilution for CUT&Tag) anti-rabbit -Actin (Cell Signaling Technology #8457, 1:1000 dilution for Western Blot) anti-mouse CEBP (Thermo Fisher #MA1-827, 1:1000 dilution for Western Blot; 1:50 dilution for CUT&Tag) anti-rabbit CEBP (Thermo Fisher #PA5-80468, 1:1000 dilution for Western Blot; 1:50 dilution for CUT&Tag) anti-mouse CHOP (Cell Signaling Technology #2895T, 1:1000 dilution for Western Blot; 1:50 dilution for CUT&Tag). anti-rabbit H3K27Ac (Abcam #ab4729, 1:50 dilution for CUT&Tag) anti-rabbit IgG (Beyotime #A7016, 1:200 dilution for immunouorescence) anti-rabbit IgG -Alexa Fluor 594 secondary antibody (Thermo Fisher, #A-11012, 1:1000 dilution for immunouorescence) anti-rabbit IgG-HRP secondary antibody (YEASEN #33101ES60, 1:10000 dilution for Western Blot) anti-mouse IgG-HRP secondary antibody (YEASEN #33201ES60,1:10000 dilution for Western Blot) Verification of all antibodies used in the study for species and application can be found at the manufacturer's website. .. 4 nature portfolio | rep orting sum m ary April2023 Eukaryotic cell lines Policy information about cell lines and Sex and Gender in Research Cell line source(s) Authentication Mycoplasma contamination Commonly misidentified lines (See ICLAC register) Novel plant genotypes Seed stocks Authentication Plants anti-rabbit ATF4 (https://www.cellsignal.com/products/primary-antibodies/atf-4-d4b8-rabb) anti-mouse ATF4 (https://www.scbt.com/p/atf-4-antibody-b-3) anti-rabbit -Actin (https://www.cellsignal.com/products/primary-antibodies/b-actin-d6a8-rabbit-mab/8457) anti-mouse CEBP (https://www.thermofisher.cn/cn/zh/antibody/product/C-EBP-beta-Antibody-clone-1H7-Monoclonal/MA1-827) anti-rabbit CEBP (https://www.thermofisher.cn/cn/zh/antibody/product/C-EBP-gamma-Antibody-Polyclonal/PA5-80468) anti-mouse CHOP (https://www.cellsignal.com/products/primary-antibodies/chop-l63f7-mouse-mab/2895) anti-rabbit H3K27Ac (https://www.abcam.cn/products/primary-antibodies/histone-h3-acetyl-k27-antibody-chip-grade-ab4729.html) anti-rabbit IgG (https://www.beyotime.com/product/A7016.htm) anti-rabbit IgG -Alexa Fluor 594 secondary antibody (https://www.thermofisher.cn/cn/zh/antibody/product/Goat-anti-Rabbit-IgG-HL-Cross-Adsorbed-Secondary-Antibody-Polyclonal/A-11012) anti-rabbit lgG-HRP secondary antibody (https://www.yeasen.com/products/detail/319) anti-mouse IgG-HRP secondary antibody (https://www.yeasen.com/products/detail/407) C2C12 cell line (# SCSP-505) and HeLa cell line (#SCSP-504) were purchased from the Cell Bank of the Chinese Academy of Sciences.
Immunoprecipitation:Article Title: Pre-established ATF4 occupancy and chromatin organization instruct selective transcription activation during integrated stress response
Article Snippet: .. This is a study with data collection and analyses relying on objective measures. anti-rabbit ATF4 (Cell Signaling Technology #11815; 1:1000 dilution for Western Blot; 1:200 dilution for immunouorescence; 1:50 dilution for CUT&Tag; 1:50 dilution for immunoprecipitation ) anti-mouse ATF4 (Santa Cruz Biotechnology #sc-390063,1:50 dilution for CUT&Tag) anti-rabbit -Actin (Cell Signaling Technology #8457, 1:1000 dilution for Western Blot) anti-mouse CEBP (Thermo Fisher #MA1-827, 1:1000 dilution for Western Blot; 1:50 dilution for CUT&Tag) anti-rabbit CEBP (Thermo Fisher #PA5-80468, 1:1000 dilution for Western Blot; 1:50 dilution for CUT&Tag) anti-mouse CHOP (Cell Signaling Technology #2895T, 1:1000 dilution for Western Blot; 1:50 dilution for CUT&Tag). anti-rabbit H3K27Ac (Abcam #ab4729, 1:50 dilution for CUT&Tag) anti-rabbit IgG (Beyotime #A7016, 1:200 dilution for immunouorescence) anti-rabbit IgG -Alexa Fluor 594 secondary antibody (Thermo Fisher, #A-11012, 1:1000 dilution for immunouorescence) anti-rabbit IgG-HRP secondary antibody (YEASEN #33101ES60, 1:10000 dilution for Western Blot) anti-mouse IgG-HRP secondary antibody (YEASEN #33201ES60,1:10000 dilution for Western Blot) Verification of all antibodies used in the study for species and application can be found at the manufacturer's website. .. 4 nature portfolio | rep orting sum m ary April2023 Eukaryotic cell lines Policy information about cell lines and Sex and Gender in Research Cell line source(s) Authentication Mycoplasma contamination Commonly misidentified lines (See ICLAC register) Novel plant genotypes Seed stocks Authentication Plants anti-rabbit ATF4 (https://www.cellsignal.com/products/primary-antibodies/atf-4-d4b8-rabb) anti-mouse ATF4 (https://www.scbt.com/p/atf-4-antibody-b-3) anti-rabbit -Actin (https://www.cellsignal.com/products/primary-antibodies/b-actin-d6a8-rabbit-mab/8457) anti-mouse CEBP (https://www.thermofisher.cn/cn/zh/antibody/product/C-EBP-beta-Antibody-clone-1H7-Monoclonal/MA1-827) anti-rabbit CEBP (https://www.thermofisher.cn/cn/zh/antibody/product/C-EBP-gamma-Antibody-Polyclonal/PA5-80468) anti-mouse CHOP (https://www.cellsignal.com/products/primary-antibodies/chop-l63f7-mouse-mab/2895) anti-rabbit H3K27Ac (https://www.abcam.cn/products/primary-antibodies/histone-h3-acetyl-k27-antibody-chip-grade-ab4729.html) anti-rabbit IgG (https://www.beyotime.com/product/A7016.htm) anti-rabbit IgG -Alexa Fluor 594 secondary antibody (https://www.thermofisher.cn/cn/zh/antibody/product/Goat-anti-Rabbit-IgG-HL-Cross-Adsorbed-Secondary-Antibody-Polyclonal/A-11012) anti-rabbit lgG-HRP secondary antibody (https://www.yeasen.com/products/detail/319) anti-mouse IgG-HRP secondary antibody (https://www.yeasen.com/products/detail/407) C2C12 cell line (# SCSP-505) and HeLa cell line (#SCSP-504) were purchased from the Cell Bank of the Chinese Academy of Sciences.
other:
Membrane:Article Title: ER Stress is Involved in Epithelial-To-Mesenchymal Transition of Alveolar Epithelial Cells Exposed to a Hypoxic Microenvironment
Article Snippet: Total protein content in lung homogenates or in AECs was determined by BCA assay (Thermo Fisher, Illkirch cedex, France). .. Samples (30 μg protein per sample in Laemmli buffer containing 5% β-mercapto-ethanol) were then resolved through 7.5% or 12% acrylamide gels, electrically transferred to nitrocellulose membrane, and incubated overnight at 4 °C with 1:2000 anti-mouse ATF4 (sc-200, Santa Cruz Biotechnology, Cliniscience, Nanterre, France) anti-mouse α-SMA (A2547, Sigma Aldrich, Saint Quentin Fallavier, France), anti-mouse vimentin (V6630, Sigma Aldrich, Saint Quentin Fallavier, France) or with 1:500 anti-rabbit ATF6α (sc-22799, Santa Cruz Biotechnology, Cliniscience, Nanterre, France), anti-mouse XBP1 (sc-7160, Santa Cruz Biotechnology, Nanterre, France), anti-rabbit HIF-1α (NB100-479, Novus, Biotechne, Lille, France) antibody solution. .. Next day, the membrane was incubated with appropriate anti-rabbit or mouse HRP-conjugated secondary antibody (Dako, P0448 and P0447 respectively, DAKO, Agilent technologies, Les Ulis Cedex, France).
Incubation:Article Title: ER Stress is Involved in Epithelial-To-Mesenchymal Transition of Alveolar Epithelial Cells Exposed to a Hypoxic Microenvironment
Article Snippet: Total protein content in lung homogenates or in AECs was determined by BCA assay (Thermo Fisher, Illkirch cedex, France). .. Samples (30 μg protein per sample in Laemmli buffer containing 5% β-mercapto-ethanol) were then resolved through 7.5% or 12% acrylamide gels, electrically transferred to nitrocellulose membrane, and incubated overnight at 4 °C with 1:2000 anti-mouse ATF4 (sc-200, Santa Cruz Biotechnology, Cliniscience, Nanterre, France) anti-mouse α-SMA (A2547, Sigma Aldrich, Saint Quentin Fallavier, France), anti-mouse vimentin (V6630, Sigma Aldrich, Saint Quentin Fallavier, France) or with 1:500 anti-rabbit ATF6α (sc-22799, Santa Cruz Biotechnology, Cliniscience, Nanterre, France), anti-mouse XBP1 (sc-7160, Santa Cruz Biotechnology, Nanterre, France), anti-rabbit HIF-1α (NB100-479, Novus, Biotechne, Lille, France) antibody solution. .. Next day, the membrane was incubated with appropriate anti-rabbit or mouse HRP-conjugated secondary antibody (Dako, P0448 and P0447 respectively, DAKO, Agilent technologies, Les Ulis Cedex, France).
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