For chromatin immunoprecipiation of ATRX cells were fixed with 2mM EGS (Pierce 26103) for 45 min at room temperature in PBS. Formaldehyde was then added to 1% for 20 minutes and quenched with 125mM glycine. Chromatin was sonicated to under 500bp and lysates were immunoprecipitated using 20 μg anti-ATRX antibody (Santa Cruz sc-15408) antibody. DNA was precipitated with 20 μg carrier glycogen. Histone H3.3 chromatin immunoprecipitations were performed using Millipore chromatin Immunoprecipitation Assay Kit (17-295) as per manufacturer’s instructions and 10 μg anti-histone H3.3 antibody (Millipore 09-838) 45 (link). Telomere binding was assessed by slot blotting and probing with a 32P-labelled TTAGGG probe. Binding at other loci was assessed by Q-PCR; primers are shown in Supplementary Table 1 . Quantification was performed using ImageJ software and presented as a percentage of the input DNA.
Anti histone h3.3 antibody
Manufactured by Merck Group
The Anti-histone H3.3 antibody is a laboratory research tool used to detect and study the histone H3.3 protein. Histones are a group of proteins that play a crucial role in the organization and regulation of DNA within eukaryotic cells. The Anti-histone H3.3 antibody specifically recognizes the H3.3 variant of the histone H3 protein, which is associated with active gene transcription and a range of cellular processes.
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Lab products found in correlation
2 protocols using anti histone h3.3 antibody
1
Chromatin Immunoprecipitation for ATRX and H3.3
2
ChIP-seq Protocol for ATRX and H3.3
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For ChIP of ATRX cells were fixed with 2 mM ethylene glycol bis(succinimidyl succinate) (EGS) (Pierce 26103) for 45 min at room temperature in PBS. Formaldehyde was then added to 1% for 20 min and quenched with 125 mM glycine. Chromatin was sonicated to under 500 bp and lysates were immunoprecipitated using 20 μg anti-ATRX antibody (Santa Cruz sc-15408) antibody. DNA was precipitated with 20 μg carrier glycogen. Histone H3.3 chromatin immunoprecipitations were performed using Millipore chromatin Immunoprecipitation Assay Kit (17-295) as per manufacturer's instructions and 10 μg anti-histone H3.3 antibody (Millipore 09-838)45 (link). Telomere binding was assessed by slot blotting and probing with a 32P-labelled TTAGGG probe. Binding at other loci was assessed by quantitative-PCR (qPCR); primers are shown in Supplementary Table 1 . Quantification was performed using ImageJ software and presented as a percentage of the input DNA.
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