Chromatin was extracted from naive CD4+Foxp3− T cells polarized for 48–72 h under various conditions (1×106 cells) after fixation with formaldehyde. Anti-H3K9me3 (61013; Active Motif), anti-dimethyl-H3 (diMe-Lys9) (D5567; Sigma-Aldrich), anti-H3K27me2 (ab24684; Abcam), anti-H3K4me3 (17-678; Millipore), anti-H3K36me3 (17-10493; Millipore), anti-H3K79me3 (17-10130; Millipore), anti-RORγt (sc-28559; Santa Cruz) and isotype-matched control antibody (sc-2027; Santa Cruz) were used for the immunoprecipitation of chromatin with an EZ ChIP kit according to the manufacturer’s instructions (Millipore). For cells transduced with retrovirus expressing FLAG-RelB, anti-FLAG (F1804; Sigma-Aldrich) was used. The precipitated DNA was then analyzed by real-time PCR. Data are presented as relative binding based on normalization to input DNA (Xiao et al., 2015 ). The sequences of all primers used in this study are given in Table S2 .
Anti h3k79me3
Manufactured by Merck Group
Anti-H3K79me3 is a laboratory reagent used to detect the presence of the trimethylated form of histone H3 lysine 79 (H3K79me3) in biological samples. It is a specific antibody that binds to this epigenetic modification, which is associated with transcriptional regulation and chromatin organization.
Automatically generated - may contain errors
Lab products found in correlation
Anti flag, by Merck Group (2 mentions)
Ab24684, by Abcam (1 mentions)
Anti h3k4me3, by Merck Group (1 mentions)
Ez chip kit, by Merck Group (1 mentions)
Anti h3k36me3, by Merck Group (1 mentions)
Sc 2027, by Santa Cruz Biotechnology (1 mentions)
Anti h3k27me2, by Abcam (1 mentions)
Anti h3k9me3, by Active Motif (1 mentions)
Anti h3k9ac, by Merck Group (1 mentions)
Anti snail, by Cell Signaling Technology (1 mentions)
2 protocols using anti h3k79me3
1
Chromatin Immunoprecipitation Assay of T Cell Epigenetics
2
ChIP Assay for Transcriptional Regulators
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The ChIP assay was performed according to routine operation. Rabbit monoclonal anti-Snail (1:500; Cell Signaling), anti-FLAG (1:2000, Sigma), rabbit monoclonal anti-H3K9ac (1:500; Millipore), and rabbit monoclonal anti-H3K79me3 (1:500; Millipore) antibodies were used in the ChIP assays with a rabbit monoclonal immunoglobulin G (IgG) (1:500; Cell Signaling) as a negative control. The presence of binding regions detected by the indicated antibodies was assessed by PCR. A small amount of precleared DNA (before the addition of the antibodies) was set as an input control and used to normalize the ChIP enriched DNA. The PCR primer sequences of the DNA fragments as parts of the targeted promoters are provided in Table S1 . The ChIP signal from the control groups was set to 1.00. The other groups were relative to the corresponding controls.
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