MDCK cells, plated on collagen I-coated PDMS, were fixed in 4% (v/v) paraformaldehyde for 15 min, permeabilized in 0.5% (v/v) Triton X-100 for 7 min, and blocked in PBS containing 0.2% (w/v) BSA and 1% goat or donkey serum at room temperature for 60 min. Primary antibodies used for immunofluorescence staining were: β-catenin (610154; BD Biosciences, San Jose, CA), phospho-γH2A.X (ab11174; Abcam), p53 (92825; Cell Signaling Technology, Danvers, MA), p53BP1 (sc-10915; Santa Cruz Biotechnology, Dallas, TX), and pY654 β-catenin/mouse (Developmental Studies Hybridoma Bank). EdU incorporation was performed using a Click-iT Plus EdU Alexa Fluor 647 Imaging Kit (C10339; Molecular Probes/Invitrogen, Waltham, MA) as directed by the manufacturer.
Phospho γh2a x
Manufactured by Abcam
Phospho-γH2A.X is a lab equipment product that detects the phosphorylation of histone H2A.X, a marker of DNA double-strand breaks. It provides a reliable and sensitive means to measure this cellular response to DNA damage.
Automatically generated - may contain errors
Lab products found in correlation
Click it plus edu alexa fluor 647 imaging kit, by Thermo Fisher Scientific (1 mentions)
β catenin, by BD (1 mentions)
P53bp1, by Santa Cruz Biotechnology (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Paraformaldehyde (pfa), by Santa Cruz Biotechnology (1 mentions)
Acetylated alpha tubulin, by Cell Signaling Technology (1 mentions)
Alpha tubulin, by LI COR (1 mentions)
β tubulin, by Cell Signaling Technology (1 mentions)
2 protocols using phospho γh2a x
1
Immunofluorescence Staining of MDCK Cells
2
Multiparameter Immunofluorescence Staining Protocol
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Immunofluorescence staining was performed according to standard protocols as described previously (Al-Khadairi et al., 2019) . Briefly, 80,000 cells were plated on Poly-Lysine coated glass coverslips (Corning), fixed with 4% paraformaldehyde (ChemCruz) and permeabilized with 0.1% Triton X-100 (Sigma). Primary antibodies against β-tubulin (Cell Signaling), phospho-γH2AX (Abcam), alpha-tubulin (Li-Cor), acetylated alpha-tubulin (Cell Signaling), MAP1B
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