Cells were plated in a 96-well glass bottom dish. Following the indicated treatments, cells were pre-extracted in ice-cold pre-extraction buffer (20 mM HEPES, 50 mM NaCl, 3 mM MgCl2, 300 mM sucrose, 0.5% Triton X-100), and subsequently fixed with 3% paraformaldehyde, 2% sucrose. Cells were then permeabilized (20 mM HEPES, 50 mM NaCl, 3 mM MgCl2, 300 mM sucrose, 0.5% Triton X-100), and blocked (1 mg/ml BSA, 5% Goat Serum, 0.1% Triton X-100, 1mM EDTA). EdU incorporation was detected by a click reaction using Alexa Fluor 488 azide (Invitrogen). The proximity ligation assay was then performed as per manufacturer’s instructions (Sigma) using a HMCES polyclonal antibody (Sigma HPA044968). Images were acquired using the 20X objective on an ImageXpress (Molecular Devices) and analyzed with MetaXpress software.
Hmces polyclonal antibody
Manufactured by Merck Group
The HMCES polyclonal antibody is a laboratory reagent used for the detection and analysis of the HMCES (5-hydroxymethylcytosine (5hmC) binding, ES cell-specific) protein in various biological samples. The antibody is produced by immunizing animals with a specific antigen derived from the HMCES protein, allowing for the identification and quantification of the target protein in experimental procedures.
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2 protocols using hmces polyclonal antibody
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2
Quantifying DNA Damage Response with PLA
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Cells were plated in a 96-well glass bottom dish. Following the indicated treatments, cells were pre-extracted in ice-cold pre-extraction buffer (20 mM HEPES, 50 mM NaCl, 3 mM MgCl2, 300 mM sucrose, 0.5% Triton X-100), and subsequently fixed with 3% paraformaldehyde, 2% sucrose. Cells were then permeabilized (20 mM HEPES, 50 mM NaCl, 3 mM MgCl2, 300 mM sucrose, 0.5% Triton X-100), and blocked (1 mg/ml BSA, 5% Goat Serum, 0.1% Triton X-100, 1mM EDTA). EdU incorporation was detected by a click reaction using Alexa Fluor 488 azide (Invitrogen). The proximity ligation assay was then performed as per manufacturer’s instructions (Sigma) using a HMCES polyclonal antibody (Sigma HPA044968). Images were acquired using the 20X objective on an ImageXpress (Molecular Devices) and analyzed with MetaXpress software.
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