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α h3k27me3

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α-H3K27me3 is a laboratory reagent used for the detection and quantification of histone H3 trimethylated at lysine 27 (H3K27me3). It is a highly specific antibody that can be used in various applications, such as Western blotting, immunohistochemistry, and chromatin immunoprecipitation (ChIP) assays, to study epigenetic modifications and gene regulation.

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Lab products found in correlation

αh3k4me3, by Abcam (2 mentions) Nextseq 500, by Illumina (1 mentions) α5hmc, by Active Motif (1 mentions) β actin, by Abcam (1 mentions) α h3k4me1, by Abcam (1 mentions) Cell lysis buffer, by Cell Signaling Technology (1 mentions) Protease inhibitor cocktail, by Cell Signaling Technology (1 mentions) α gapdh, by Santa Cruz Biotechnology (1 mentions) Phenyl methyl sulfonyl fluoride (pmsf), by Merck Group (1 mentions) Dual color protein loading buffer, by Thermo Fisher Scientific (1 mentions) Restore western blot stripping buffer, by Thermo Fisher Scientific (1 mentions) Pvdf membrane, by GE Healthcare (1 mentions)

5 protocols using α h3k27me3

1

Multiplexed ChIP-seq Protocol

Cited 1 time
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Multiplexed Chromatin immunoprecipitation followed by next generation sequencing (MINT-ChIP-seq) was performed as previously described (41 (link)) using the following antibodies: α-HA (ab9110), α-H3 (Active-Motif), α-H3K27me3 (Millipore, 07-449) and αH3K27ac (a gift from Hitoshi Kimura, Osaka University). Sequencing was performed in-house using Illumina NextSeq500.
Cohen L.R., Kaffe B., Deri E., Leibson C., Nissim-Rafinia M., Maman M., Harpaz N., Ron G., Shema E, & Meshorer E. (2022). PRC2-independent actions of H3.3K27M in embryonic stem cell differentiation. Nucleic Acids Research, 51(4), 1662-1673.
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2

Comprehensive Protein Expression Analysis

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α-L2HGDH (Proteintech, #15707–1-AP, GeneTex, #GTX32695), α−5hmc (Active Motif, #39791), α-KDM6A (GeneTex, #GTX120873), α-MDH2 (Abcam, #ab96193), α-H3K27Me3 (Millipore, #07–449), α-H3 (CST, #9715), α-EZH2 (CST, #5246) and β-actin (Abcam, #ab20272) were used as per manufacturers’ instructions.
Shelar S.B., Shim E.H., Brinkley G.J., Kundu A., Carobbio F., Poston T., Tan J., Parekh V., Benson D., Crossman D.K., Buckhaults P.J., Rakheja D., Kirkman R., Sato Y., Ogawa S., Dutta S., Velu S.E., Emberley E., Pan A., Chen J., Huang T., Absher D., Becker A., Kunick C, & Sudarshan S. (2018). Biochemical and Epigenetic Insights into L-2-Hydroxyglutarate, a Potential Therapeutic Target in Renal Cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 24(24), 6433-6446.
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3

Chromatin Modification and Transcription Analysis

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α-H3K4me3 (Abcam[UK]-8580), α-H3K4me1(Abcam-8895), α-H3K27me3 (Millipore[UK]-07-449), α-H3K9/K14ac (Abcam 12,179), α-H3 (Abcam 1791), α-SUZ12 (Abcam 12,073-100), α-RNA Pol II N20 (Santa-Cruz[UK] 899), α-RNA Pol II S5P (Abcam 5131), α-RNA Pol II unphosphorylated CTD (Abcam 817), α-GFP (Chromotec [Germany] GFP-TRAP-A gta-20), α-IgG (Invitrogen 10500C).
Wachter E., Quante T., Merusi C., Arczewska A., Stewart F., Webb S, & Bird A. (2014). Synthetic CpG islands reveal DNA sequence determinants of chromatin structure. eLife, 3, e03397.
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4

Western Blot Analysis of Histone Modifications

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Cells were grown in 10-cm2 dishes and lysed using Cell Lysis Buffer (Cell Signaling) with protease inhibitor cocktail (Cell Signaling) and 100 µM PMSF (SIGMA). Protein samples were resolved on 12 % Tris–glycine gels and transferred to PVDF membranes. The membranes were then incubated with α-H3K27me3 (1:1000; Millipore, 07-449), α-H3 (1:1000; SIGMA, H0164), α-BAK1 (Santa Cruz, sc-832) and α-GAPDH (Santa Cruz, sc-25778). After washing with TBS, the membranes were incubated with peroxidase-conjugated goat anti-rabbit antibody (1:20,000; Novus, NB7187) followed by chemiluminescence staining (Millipore).
Cisneros-Soberanis F., Andonegui M.A, & Herrera L.A. (2016). miR-125b-1 is repressed by histone modifications in breast cancer cell lines. SpringerPlus, 5(1), 959.
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5

Chromatin Modifications in Drosophila Testes

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We dissected testes from bam1/bam114–97 and E(z)61 bam1/E(z)61 bam114–97 flies, which were raised at 29°C for 3 days. Samples were grouped in DualColor Protein Loading Buffer (Fermentas, #R1011) and boiled for 10 mins, followed by electrophoresis using a 12% SDS-PAGE gel and transfer to a PVDF membrane (GE healthcare, #RPN303F). The membrane was then blotted with α-H3K27me3 (Millipore, #07-449) at 1:200 overnight at 4°C, followed by HRP-conjugated α-rabbit IgG at 1:1,000 (Invitrogen, # 656120) at RT for 2 hrs. The ECL (GE healthcare, RPN2109) reagents were applied to visualize the signal. After repeated stripping with Restore Western Blot Stripping buffer (Thermo scientific, #21059), the membrane was re-probed sequentially with α-H3K4me3 (Abcam, #ab8580) at 1:10,000 and α-H3 (Abcam, #ab1791) at 1: 5000, followed by HRP-conjugated rabbit IgG at 1:1,000.
Eun S.H., Feng L., Cedeno-Rosario L., Gan Q., Wei G., Cui K., Zhao K, & Chen X. (2017). Polycomb group gene E(z) is required for spermatogonial dedifferentiation in Drosophila adult testis. Journal of molecular biology, 429(13), 2030-2041.
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