Antibodies used in this study: anti‐ASH2L (A300‐107A; Bethyl Laboratories), anti‐ASH2L (12331‐1‐AP; Proteintech Group), anti‐FLAG (4110‐FG; GNI), anti‐ERα (D8H8) (#8664; Cell Signaling Technology), anti‐ERα (F10) (sc‐8002; Santa Cruz Biotechnology), anti‐MLL1 (A300‐37A; Bethyl Laboratories), anti‐WDR5 (A302‐429A; Bethyl Laboratories), anti‐PAX2 (TA327502S; OriGene Technologies), anti‐Cyclin D1 (60186‐1‐lg; Proteintech Group), anti‐GAPDH (AC033; ABclonal Technology), anti‐Ki67 (sc‐15402; Santa Cruz Biotechnology), anti‐trimethyl H3‐K27 (07‐449; Millipore), anti‐trimethyl H3‐K4 (05‐745R; Millipore).
Anti trimethyl h3k27
Manufactured by Merck Group
Sourced in United States
Anti-trimethyl H3K27 is a laboratory antibody used to detect and quantify trimethylation of histone H3 at lysine 27 (H3K27me3), a well-known epigenetic mark associated with transcriptional repression. This antibody can be used in various applications such as chromatin immunoprecipitation (ChIP), western blotting, and immunohistochemistry to study the role of H3K27 trimethylation in biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Anti trimethyl h3k4, by Merck Group (5 mentions)
Anti mll1, by Fortis Life Sciences (1 mentions)
Anti ki67, by Santa Cruz Biotechnology (1 mentions)
Anti erα d8h8, by Cell Signaling Technology (1 mentions)
Anti erα f10, by Santa Cruz Biotechnology (1 mentions)
Anti wdr5, by Fortis Life Sciences (1 mentions)
Anti gapdh, by ABclonal (1 mentions)
Anti cyclin d1, by Proteintech (1 mentions)
Ab9110, by Abcam (1 mentions)
Anti foxa1, by Abcam (1 mentions)
Anti β tubulin, by Santa Cruz Biotechnology (1 mentions)
Anti ar, by Abcam (1 mentions)
Anti pol 2, by Santa Cruz Biotechnology (1 mentions)
Anti acetylated h3k27, by Abcam (1 mentions)
Rhodamine phalloidin, by Merck Group (1 mentions)
Anti n cadherin, by Merck Group (1 mentions)
Anti apkc, by Santa Cruz Biotechnology (1 mentions)
Anti caspase 3, by BD (1 mentions)
Anti β tubulin, by Merck Group (1 mentions)
Anti mouse igg hrp, by GE Healthcare (1 mentions)
8 protocols using anti trimethyl h3k27
1
Antibody Panel for Cell Signaling Analysis
2
Chromatin Immunoprecipitation Assay for Arabidopsis
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For the ChIP assay, Col-0, pGI:GI-HA gi-2, and pGI:GI-HA gi-2
det1-1 plants were grown for 10 days under SD (8 h light:16 h dark)
conditions and collected at ZT8. The samples were cross-linked with 1%
formaldehyde, ground to powder in liquid nitrogen, and then sonicated43 (link). The sonicated chromatin complexes were bound with anti-HA
antibody (ab9110, Abcam) for immunoprecipitation. The amount of DNA fragment was
analyzed by quantitative real-time PCR (qPCR) using specific primers.
UBI10 was used as an internal standard for normalization. The primers
used for qPCR are listed inTable S2 . For another ChIP
assay, Col-0 and det1-1 plants were grown for 14 days under SD (8 h
light/16 h dark) conditions and collected at ZT8. For immunoprecipitation, we
used the anti-trimethyl H3K4 (07-473, Millipore), and anti-trimethyl H3K27
(07-449, Millipore). FUS3 was used as an internal standard for
normalization14 (link). Experiments were performed with three
biological repeats.
det1-1 plants were grown for 10 days under SD (8 h light:16 h dark)
conditions and collected at ZT8. The samples were cross-linked with 1%
formaldehyde, ground to powder in liquid nitrogen, and then sonicated43 (link). The sonicated chromatin complexes were bound with anti-HA
antibody (ab9110, Abcam) for immunoprecipitation. The amount of DNA fragment was
analyzed by quantitative real-time PCR (qPCR) using specific primers.
UBI10 was used as an internal standard for normalization. The primers
used for qPCR are listed in
assay, Col-0 and det1-1 plants were grown for 14 days under SD (8 h
light/16 h dark) conditions and collected at ZT8. For immunoprecipitation, we
used the anti-trimethyl H3K4 (07-473, Millipore), and anti-trimethyl H3K27
(07-449, Millipore). FUS3 was used as an internal standard for
normalization14 (link). Experiments were performed with three
biological repeats.
3
Prostate Cancer Cell Line Maintenance and Antibodies
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Human prostate cancer LNCaP and C4-2B cells were described previously [25 (link)]. All cell lines were maintained in RPMI 1640 supplemented with 10% (v/v) fetal bovine serum (FBS). Antibodies are: anti-AR (Abcam, ab74272), anti-FOXA1 (Abcam, ab23738), anti-acetylated H3K9/14 (Millipore, #06–599), anti-acetylated H3K27 (Abcam, ab4729), ant-dimethyl-H3K4 (Millipore, #07–030), ant-trimethyl-H3K4 (Millipore, #07–473), anti-trimethyl-H3K27 (Millipore, #07–449), anti-Pol II (Santa Cruz Technology, sc-899), and anti-β-tubulin (Santa Cruz Technology, sc-80011).
4
Immunolabeling of Neural Stem Cells
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The following antibodies were used: anti-BrdU (Hybridoma Bank); anti-mouse β-tubulin III (Tuj1, Covance, MMS-435P); anti-H3S10p (Upstate); anti-caspase 3 (BD PharMingen); anti-N-cadherin (DSHB); anti-trimethyl H3K27 (Millipore); anti-EZH2 (Zymed and Active motif); anti-aPKC (Santa Cruz, sc-17781); anti-β-tubulin (Millipore, MAB3408); anti-SOX5 (kindly provided by Dr Morales [55 (link)]; anti-pY31 paxillin (Invitrogen), Actin was stained with phalloidin-rhodamine (Sigma, P1951).
5
Histone Modification Analysis Protocol
6
Chromatin Immunoprecipitation (ChIP) Assay Protocol
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Chromatin Immunoprecipitation (ChIP) assays were performed essentially as described before (Coarfa et al., 2020 (link); Hu et al., 2020 (link); Jehanno et al., 2020 (link); Maity et al., 2020 (link); Mallik et al., 2020 (link); Moon et al., 2020 (link); Shen et al., 2020 (link); Wang J. N. et al., 2020 (link); Wang S. et al., 2020 (link); Zhang et al., 2020 (link); Zhao et al., 2020 (link); Marti et al., 2021 (link); Peng et al., 2021 (link); Rashid et al., 2021 (link)). In brief, chromatin in control and treated cells were cross-linked with 1% formaldehyde. Cells were incubated in lysis buffer (150 mM NaCl, 25 mM Tris pH 7.5, 1% Triton X-100, 0.1% SDS, 0.5% deoxycholate) supplemented with protease inhibitor tablet and PMSF. DNA was fragmented into ∼200 bp pieces using a Branson 250 sonicator. Aliquots of lysates containing 200 μg of protein were used for each immunoprecipitation reaction with anti-acetyl H3 (Millipore, 06-599), anti-trimethyl H3K4 (Millipore, 07-473), anti-trimethyl H3K9 (Diagenode, C15410193), anti-trimethyl H3K27 (Millipore, 04-449), anti-trimethyl H4K20 (Millipore, 07-463), anti-KDM4A (Abcam, ab191433), anti-KDM4B (Bethyl Laboratories, A301-477A), anti-KDM4C (Bethyl Laboratories, A300-885A), or anti-KDM4D (Proteintech, 22591-1-AP) antibodies.
7
Chromatin Immunoprecipitation of Histone Modifications
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Chromatin immunoprecipitation (ChIP) assays were performed on 2-week-old seedlings according to a previously described method (Saleh et al., 2008 (link)) using the following antibodies: anti-trimethyl-H3K27 (07-449; Millipore), anti-trimethyl-H3K4 (07-473; Millipore), and anti-dimethyl-H3K9 (ab1220; Abcam). The gene-specific primers used in PCR are listed in Supplementary Table S1 .
8
Chromatin Immunoprecipitation Assays for Seed Epigenetics
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ChIP assays were conducted following previously described procedures with minor modifications (Wang et al., 2014) . Total chromatin extracted from seeds was subjected to ChIPs with anti-Flag (F7425, Sigma), anti-HA (H6908, Sigma), anti-trimethyl H3K27 (07-449; Millipore, Burlington, MA, USA), or anti-trimethyl H3K36 (07-549; Millipore). Genomic fragments of FLC and TUB2 (internal control gene) were measured by qPCR in an ABI QuantStudio 5/6 Flex system with an SYBR Green PCR master mix. To calculate relative fold changes of ABI3:Flag, FRI:HA and Flag:ABI5 on FLC chromatin, the level of a particular FLC fragment was first normalized by that of the internal control TUB2, and then further normalized by the level of this fragment in a background control sample (CK). Three biological replicates were conducted in each ChIP assay; qPCR primers are described in Supplemental Table S1.
For the analysis of ABI3:Flag enrichment in the absence of LEC1 function, an ABI3:Flag (in lec1 FRI) line was crossed to WT (FRI-Col) and lec1 FRI, and 6-DAP F 1 seeds were harvested for ChIP.
For the analysis of ABI3:Flag enrichment in the absence of LEC1 function, an ABI3:Flag (in lec1 FRI) line was crossed to WT (FRI-Col) and lec1 FRI, and 6-DAP F 1 seeds were harvested for ChIP.
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