ChIP-seq-grade antibodies directed against histone modifications were purchased from Diagenode: H3K9ac C15410004, H3K27me3 C15410195, and H3K4me3 C15410003. For the antibody against P. tetraurelia RPB1, the peptide SPHYTSHTNSPSPSYRSS-C was used for immunization. Purification and testing of specificity by western blots and immunostaining were performed as described recently (Drews et al. 2021 (link)). Because there are some amino acid differences in the N-terminal tail of the Paramecium H3P1 to Human H3 (Supplemental Fig. S1A ), the peptide PtH3K27me3 TKAARK(me3)TAPAVG was synthesized, and binding affinity of the purchased H3K27me3 antibody to the PtH3k27me3 peptide was verified by dot-blots and competition assays. For details, see Supplemental Methods .
H3k27me3 c15410195
Manufactured by Diagenode
The H3K27me3 C15410195 is a specific antibody that recognizes the trimethylated form of histone H3 at lysine 27 (H3K27me3). This epigenetic mark is associated with transcriptional repression and is involved in the regulation of gene expression.
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2 protocols using h3k27me3 c15410195
1
Antibody Validation for Histone Modifications
2
Validation of Histone Modification Antibodies
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Polyclonal, ChIP-seq grade antibodies directed against histone modifications were purchased from Diagenode: H3K9ac C15410004, H3K27me3 C15410195, H3K4me3 C15410003. For antibody against P. tetraurelia RBP1, the peptide SPHYTSHTNSPSPSYRSS-C was used for rabbit immunisation. Purification and testing of specificity by Western blots and immunostaining was carried out as described recently [18] . Since there are some amino acid differences in the N-terminal tail of the Paramecium H3P1 to Human H3 (Suppl. Fig. 1A), the peptide PtH3K27me3 TKAARK(me3)TAPAVG 3/27 was synthesized and binding affinity of the purchased H3K27me3 antibody to the PtH3k27me3 peptide was verified by dotblots and competition assays. Peptide competition assays (Suppl.Fig. 1B) were performed by blocking 2 µg of each antibody with a 10 fold excess of its corresponding peptide over night at 4 • C with agitation. 1 pmol to 100 pmol of each peptide were blotted on a nitrocellulose membrane and decorated with blocked and unblocked antibodies.
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