Cells were labeled with 100 µM iododeoxyuridine (IdU) for 10 min and then labeled with100 µM chlorodeoxyuridine (CldU) for 20 min for mock experiment. For monitoring DNA synthesis during replication stress, cells were labeled with IdU (100 µM) for 10 min, followed by exposure to CldU (100 µM) coupled with hydroxyurea (5 mM for HCT115 and 0.5 mM for human fibroblasts) for 1 h. DNA fibers were spread as described previously (29 (link)) and stained with primary antibodies (mouse anti-BrdU/IdU from BD Bioscience; rat anti-BrdU/CldU from Accurate Chemical) and fluorescence-conjugated secondary antibodies (Alexa Fluor 488-anti-rat and Texas Red-conjugated anti-mouse from Invitrogen). Fibers were imaged using Zeiss Axio Imager M2 and measured using AxioVision software (x64 version 4.8.3.0).
Texas red conjugated anti mouse
Manufactured by Thermo Fisher Scientific
Sourced in United States
Texas Red-conjugated anti-mouse is a secondary antibody used in immunofluorescence and flow cytometry applications. It is designed to bind to mouse primary antibodies and emit fluorescence in the Texas Red spectrum when excited.
Automatically generated - may contain errors
4 protocols using texas red conjugated anti mouse
1
Dual Labeling of DNA Synthesis for Replication Monitoring
2
Immunofluorescence Staining of Stemness Markers
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Cells were cultured on coverslips overnight, fixed with 4% paraformaldehyde for 30 minutes, and then treated with 0.5% Triton X-100 for 15 minutes. After being blocked in 10% normal blocking serum at room temperature for 15 minutes, the slides were incubated with antibodies from CST (USA), including rabbit antibodies to PRRX1 (Cat# DF4274; Affinity, Cincinnati, OH, USA), CD133 (Cat# 64326; CST, USA) and SOX2 (Cat# 3579; CST, USA) (1:100) at 4 ℃ overnight, which was followed by PBS rinsing. Cover slips were incubated with antibodies from CST (USA), including a fluorescein isothiocyanate (FITC)-conjugated anti-rabbit (Cat# 4412) or mouse (Cat# 4408) stain and Texas Red-conjugated anti-mouse (Cat# 8890) or anti-rabbit antibodies (Cat# 8889) (1:200) for 30 minutes at room temperature, then stained with 6-diamidino-2-phenylindole (DAPI; Cat# D3571, Invitrogen, USA).
3
Dual DNA Replication Labeling Protocol
4
Dual DNA Replication Labeling Protocol
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Cells were labeled with 100 μM iododeoxyuridine (IdU) for 10 min, then with 100 μM chlorodeoxyuridine (CldU) for 20 min. DNA fibers were spread as described previously26 (link) and stained with primary antibodies (mouse anti-BrdU/IdU from BD Bioscience; rat anti-BrdU/CldU from Accurate Chemical) and fluorescence-conjugated secondary antibodies (Alexa Fluor 488-anti-rat and Texas Red-conjugated anti-mouse from Invitrogen). Fibers were imaged using Zeiss Axio Imager M2 and measured using AxioVision software (SE64 version 4.9.1).
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