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H3k9me3 17

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H3K9me3 is a histone modification that is associated with transcriptional repression. It is a methylation of the ninth lysine residue on the histone H3 protein. This modification is involved in the regulation of gene expression and chromatin structure.

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Lab products found in correlation

Protein a g magnetic beads, by Merck Group (2 mentions) Hells ab3851 and control igg ab46540, by Abcam (2 mentions) H3k27me3, by Merck Group (2 mentions) Sybr green master mix, by Thermo Fisher Scientific (2 mentions) S2 sonicator, by Covaris (2 mentions)

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H3K9me3 (74 citations)

2 protocols using h3k9me3 17

1

ChIP-Seq on Chromatin Fragments

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Nuclei were isolated by hypotonic lysis. Sonication to obtain chromatin fragments of around 500 bp was performed using a Covaris S2 sonicator, (Woburn, MA, USA). Each sample with an equal final concentration of 100 μg chromatin was incubated overnight with 3 μg of H3K9me3 (17–625), H3K27me3 (07–449), antibodies from Millipore (Billerica, MA, USA); HELLS (AB3851) and control IgG (AB46540) from Abcam, Cambridge, UK. DNA-antibody complexes were recovered by adding protein A/G magnetic beads (Millipore). After extensive washing, bound DNA fragments were eluted (1% w/v SDS, 0.1 M Na-HCO3) and analyzed by quantitative PCR using SYBR Green Master Mix (Applied Biosystems, Thermo Fisher Scientific). ChIP in FFPE tissue was performed with ChIP-IT® FFPE Active Motif kit (Carlsbad, CA, USA) following the manufacturer’s instructions. Precipitated DNA was analyzed by quantitative PCR as explained above for classical ChIP.
Samuelsson J.K., Dumbovic G., Polo C., Moreta C., Alibés A., Ruiz-Larroya T., Giménez-Bonafé P., Alonso S., Forcales S.V, & Manuel P. (2016). Helicase Lymphoid-Specific Enzyme Contributes to the Maintenance of Methylation of SST1 Pericentromeric Repeats That Are Frequently Demethylated in Colon Cancer and Associate with Genomic Damage. Epigenomes, 1(1), 2.
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2

ChIP-Seq on Chromatin Fragments

Check if the same lab product or an alternative is used in the 5 most similar protocols
Nuclei were isolated by hypotonic lysis. Sonication to obtain chromatin fragments of around 500 bp was performed using a Covaris S2 sonicator, (Woburn, MA, USA). Each sample with an equal final concentration of 100 μg chromatin was incubated overnight with 3 μg of H3K9me3 (17–625), H3K27me3 (07–449), antibodies from Millipore (Billerica, MA, USA); HELLS (AB3851) and control IgG (AB46540) from Abcam, Cambridge, UK. DNA-antibody complexes were recovered by adding protein A/G magnetic beads (Millipore). After extensive washing, bound DNA fragments were eluted (1% w/v SDS, 0.1 M Na-HCO3) and analyzed by quantitative PCR using SYBR Green Master Mix (Applied Biosystems, Thermo Fisher Scientific). ChIP in FFPE tissue was performed with ChIP-IT® FFPE Active Motif kit (Carlsbad, CA, USA) following the manufacturer’s instructions. Precipitated DNA was analyzed by quantitative PCR as explained above for classical ChIP.
Samuelsson J.K., Dumbovic G., Polo C., Moreta C., Alibés A., Ruiz-Larroya T., Giménez-Bonafé P., Alonso S., Forcales S.V, & Manuel P. (2016). Helicase Lymphoid-Specific Enzyme Contributes to the Maintenance of Methylation of SST1 Pericentromeric Repeats That Are Frequently Demethylated in Colon Cancer and Associate with Genomic Damage. Epigenomes, 1(1), 2.
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