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Anti h3k4me1

Manufactured by Active Motif
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Anti-H3K4me1 is a laboratory product that specifically detects histone H3 monomethylated at lysine 4 (H3K4me1). This modification is associated with active enhancers and is involved in the regulation of gene expression.

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Lab products found in correlation

Anti h3k27ac, by Active Motif (7 mentions) Anti h3k27me3, by Merck Group (4 mentions) Anti h3k9ac, by Active Motif (3 mentions) Anti h3k4me2, by Active Motif (3 mentions) Anti h3k4me3, by Active Motif (2 mentions) Anti h3, by Abcam (2 mentions) Anti h3k27me3, by Cell Signaling Technology (2 mentions) Anti h3k9me2, by ABclonal (2 mentions) Anti h3k4me3, by ABclonal (2 mentions) Anti h3k9me3, by ABclonal (2 mentions) Anti h3k27me3, by Active Motif (2 mentions) Anti p300, by Santa Cruz Biotechnology (2 mentions) Anti α tubulin, by Santa Cruz Biotechnology (1 mentions) Anti set7 9, by Cell Signaling Technology (1 mentions) α tubulin, by Santa Cruz Biotechnology (1 mentions) Anti flag, by Merck Group (1 mentions) Anti h3k4ac, by Active Motif (1 mentions) Anti h3k4me3, by Abcam (1 mentions) Anti h3k9me3, by Abcam (1 mentions) Anti h3k14ac, by Active Motif (1 mentions)

12 protocols using anti h3k4me1

1

Western Blot Immunodetection Protocol

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Western blot (WB) was performed as described previously [42 (link)]. The primary antibodies used in this study were anti-SET7/9 (rabbit monoclonal, C24B1, 1:1000; Cell Signaling Technology, Danvers, MA; and mouse monoclonal, clone 5F2.3, 1:500; Merck Millipore), anti-SREK1IP1 (rabbit polyclonal, 1:200; GeneTex Inc, Irvine, CA), anti-H3K4me1 (No. 39298, rabbit polyclonal, 1:5000; Active Motif, Carlsbad, CA), anti-FLAG (mouse monoclonal M2, 1:10,000; Sigma-Aldrich Japan), and anti-α-Tubulin (mouse monoclonal, 1:400; Santa Cruz Biotechnology, Santa Cruz, CA) antibodies.α-Tubulin was used as an internal control for WB.
Akiyama Y., Koda Y., Byeon S.J., Shimada S., Nishikawaji T., Sakamoto A., Chen Y., Kojima K., Kawano T., Eishi Y., Deng D., Kim W.H., Zhu W.G., Yuasa Y, & Tanaka S. (2015). Reduced expression of SET7/9, a histone mono-methyltransferase, is associated with gastric cancer progression. Oncotarget, 7(4), 3966-3983.
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2

Histone Modifications in Zebrafish Embryos

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Tg(cmcl2:GFP) and MZsmyd4L544Efs*1 embryos were collected at 48 hpf. Protein was obtained in RIPA containing 1 mM PMSF and complete protease inhibitors (Roche, USA) after sonication. The histone modification antibodies used in this study include anti-H3 (CST, USA), anti-H3K4ac (Active Motif, USA), anti-H3K9ac (Active Motif, USA), anti-H3K14ac (Active Motif, USA), anti-H3K27ac (Active Motif, USA), anti-H3K4me1 (Active Motif, USA), anti-H3K4me2 (Active Motif, USA), anti-H3K4me3 (Abcam, USA), anti-H3K9me3 (Abcam, USA), and anti-H3K27me3 (Millipore, USA).
Xiao D., Wang H., Hao L., Guo X., Ma X., Qian Y., Chen H., Ma J., Zhang J., Sheng W., Shou W., Huang G, & Ma D. (2018). The roles of SMYD4 in epigenetic regulation of cardiac development in zebrafish. PLoS Genetics, 14(8), e1007578.
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3

H3K4me1 ChIP-qPCR of RELA Promoter

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Chromatin from Beas-2B cells treated by scramble or siSETD7 siRNA with or without H2O2 stimulation were fixed with 37% formaldehyde. Chromatin immunoprecipitation was performed with anti-H3K4me1 (Active Motif) and ChIP-IT Express Chromatin Immunoprecipiation Kit from Active Motif, following protocols from the manufacturer. 20 ug chromatin from each treatment group and 5 μl of anti-H3K4me1 or anti-IgG antibody were used for each IP. For qPCR, primer pairs corresponding to the −250 bp ~+250 bp RELA promoter were designed. GAPDH (positive control) primer pairs and two primer pairs (negative controls) that amplify regions in gene deserts (Untr4, Untr12) were included in each test. Relative expression level of each site was shown by comparison to input chromatin of each sample.
He S., Owen D.R., Jelinsky S.A, & Lin L.L. (2015). Lysine Methyltransferase SETD7 (SET7/9) Regulates ROS Signaling through mitochondria and NFE2L2/ARE pathway. Scientific Reports, 5, 14368.
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4

ChIP-seq of histone marks and regulators

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Formaldehyde crosslinked chromatin from 5 million cells per ChIP was sheared to small fragments by Covaris M220 Focused-ultrasonicator. The following antibodies were used for chromatin pull-down in RIPA buffer with protease inhibitor cocktail: anti-H3K27ac (Active Motif 39685, 5 μg), anti-H3K4me1 (Active Motif 39297, 10 μl), anti-CTCF (Abcam ab70303, 5 μg), anti-SMC3 (Abcam ab9263, 5 μg). After capturing by Protein A/G magnetic beads, chromatin was washed 6 times and reverse-crosslinked for 3 hr with RNase A and Proteinase K. ChIP DNA library was constructed by NEBNext Ultra II DNA Library Prep kit for paired-end sequencing.
Wu S., Turner K.M., Nguyen N., Raviram R., Erb M., Santini J., Luebeck J., Rajkumar U., Diao Y., Li B., Zhang W., Jameson N., Corces M.R., Granja J.M., Chen X., Coruh C., Abnousi A., Houston J., Ye Z., Hu R., Yu M., Kim H., Law J.A., Verhaak R.G., Hu M., Furnari F.B., Chang H.Y., Ren B., Bafna V, & Mischel P.S. (2019). Circular ecDNA promotes accessible chromatin and high oncogene expression. Nature, 575(7784), 699-703.
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5

Western Blot Analysis of Yeast Lysates

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Yeast extracts were prepared using TCA lysis method as described above. Either equal amounts or a serial dilution of the lysates was prepared from various samples before resolving them in SDS-PAGE and transferring onto a polyvinylidene difluoride membrane. Following incubation with primary rabbit or mouse antibody and corresponding HRP-conjugated secondary antibody, protein signals were detected by chemiluminescence using Pierce ECL Plus Western Blotting Substrate (Thermo Scientific) and autoradiography. The following antibodies were used with their source and catalog numbers indicated within parentheses: anti-Flag M2 (F3165; Sigma), anti-Pgk1 (459250; Invitrogen), anti-GFP (AE011, Abclonal), anti-UBE2A (A7744; Abclonal), anti-UBE2B (A6315, Abclonal), anti-V5 (R690, Invitrogen), anti-H2B (39237; Active Motif), anti-H3 (ab1791; Abcam), anti-H3K4me1 (39297; Active Motif), anti-H3K4me2 (399141; Active Motif), anti-H3K4me3 (39159; Active Motif), anti-ubiquitin antibody (ab139467; Abcam); monoubiquitinylated and polyubiquitinylated conjugates monoclonal antibody (FK2) (HRP conjugate) (BML-PW0150; Enzo Life Sciences), and anti-Rad6 (DZ33919; Boster Bio). Please note that the anti-UBE2B antibody recognizes both UBE2A and UBE2B (Fig. S2).
Shukla P.K., Sinha D., Leng A.M., Bissell J.E., Thatipamula S., Ganguly R., Radmall K.S., Skalicky J.J., Shrieve D.C, & Chandrasekharan M.B. (2022). Mutations of Rad6 E2 ubiquitin-conjugating enzymes at alanine-126 in helix-3 affect ubiquitination activity and decrease enzyme stability. The Journal of Biological Chemistry, 298(11), 102524.
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6

Western Blot Analysis of Histone Modifications

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Cell lysis was incubated at 95 °C for 15 min with 1× SDS-PAGE Loading Buffer. The primary antibodies are as follows: anti-H3 (Catalog No. ab1791, Abcam, Cambridge, UK), anti-H3K9me2 (Catalog No. A2359, Abclonal, Woburn, MA), anti-H3K9me3 (Catalog No. A2360, Abclonal), anti-H3K9ac (Catalog No. 39137, Active Motif, Carlsbad, CA), anti-H3K27ac (Catalog No. 39133, Active Motif), anti-H3K27me3 (Catalog No. 9733, Cell Signaling Technology, Boston, MA), anti-H3K4me1 (Catalog No. 39297, Active Motif), and anti-H3K4me3 (Catalog No. A2357, Abclonal). HRP-conjunction secondary antibodies (Catalog No. 711-035-152, Jackson ImmunoResearch Laboratories, West Grove, PA) and Chemiluminescent HRP Substrate (Catalog No. WBKLS0500, Millipore, Billerica, MA) were used for the detection. Software AlphaView was used for the relative grayscale statistics.
Yan Z., Ji L., Huo X., Wang Q., Zhang Y, & Wen B. (2020). G9a/GLP-sensitivity of H3K9me2 Demarcates Two Types of Genomic Compartments. Genomics, Proteomics & Bioinformatics, 18(4), 359-370.
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7

Chromatin Immunoprecipitation (ChIP) Assay

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Chromatin immunoprecipitation (ChIP) assays were performed as described previously (18 (link)). Rabbit polyclonal antibodies for ChIP were either custom generated for EBNA1 (Pocono Rabbit Farm) or were purchased for rabbit and mouse anti-IgG (Santa Cruz Biotechnology), anti-HCF1 (catalog no. A301-400A; Bethyl), anti-RbBp5 (catalog no. A300-109A; Bethyl), anti-Ash2L (catalog no. A300-489A; Bethyl), anti-OCT2 (catalog no RB9297; Neo Markers) and pan-H3 (catalog no. 07-690), H3K4me3 (catalog no. 07-473), H3K9acetyl (catalog no. 07-352) were purchased from Millipore. Rabbit serum anti-H3K9me3 (catalog no. 39161), anti-H3K4me1 (catalog no. 61633), anti-H3K27me3 (catalog no. 39155)- and anti-H3K27acetyl (catalog no. 39685), and anti-H3K4me2 (catalog no. 39141) were from Active Motif.
Dheekollu J., Wiedmer A., Sentana-Lledo D., Cassel J., Messick T, & Lieberman P.M. (2016). HCF1 and OCT2 Cooperate with EBNA1 To Enhance OriP-Dependent Transcription and Episome Maintenance of Latent Epstein-Barr Virus. Journal of Virology, 90(11), 5353-5367.
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8

Chromatin Immunoprecipitation and Sequencing

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Immunoprecipitation and DNA sequencing was performed by Active Motif. The following antibodies were used: anti-H3K27me3 (Millipore 07–449), anti-p300 (Santa Cruz sc–551X), anti-H3K4me1 (Active Motif 39287), anti-H3K4me3 (Active Motive 39159), anti-H3K27ac (active Motif 39133), anti-RARα (Diagenode C15310155). Illumina sequencing libraries were prepared from the ChIP and Input DNAs. For ChIP q-PCR, enrichment calculated as binding events per 1,000 cells using Active Motif’s normalization scheme. Detailed methods for ChIP-seq and binding site analyses are provided in the Supplemental Information.
Brown C.C., Esterhazy D., Sarde A., London M., Pullabhatla V., Osma-Garcia I., al-Bader R., Ortiz C., Elgueta R., Arno M., de Rinaldis E., Mucida D., Lord G.M, & Noelle R.J. (2015). Retinoic Acid Is Essential for Th1 Cell Lineage Stability and Prevents Transition to a Th17 Cell Program. Immunity, 42(3), 499-511.
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9

Chromatin Immunoprecipitation (ChIP) Assay

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In vitro–cultured cells were cross-linked by 1% paraformaldehyde for 10 min at room temperature with gently shaking and were stopped by 100 mM glycine. The ChIP experiment was performed following the instructions of ChIP-IT Express ChIP kits (Active Motif). Antibodies, including anti-rabbit IgG [2729, Cell Signaling Technology (CST)], anti-HA (3724, CST), anti-H3K4me3 (07-473, Millipore), anti-H3K27me3 (07-499, Millipore), anti-H3K27Ac (39133, Active Motif), anti-H3K4me1 (39297, Active Motif), and Dynabeads protein A and G (Life Technologies) were used for IPs. The precipitated DNA was quantified by real-time PCR and normalized on the basis of total input DNA.
Chi X., Jin W., Zhao X., Xie T., Shao J., Bai X., Jiang Y., Wang X, & Dong C. (2022). RORγt expression in mature TH17 cells safeguards their lineage specification by inhibiting conversion to TH2 cells. Science Advances, 8(34), eabn7774.
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10

Antibody Characterization for Epigenetic Regulation

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The following antibodies were utilized in for the present study: anti-H2Aub1 (Cell Signaling Technology Cat# 8240; RRID:AB_10891618), anti-Rnf2 (Cell Signaling Technology Cat# 5694; RRID:AB_10705604), anti-H2A.Z (Active Motif Cat# 39113; RRID:AB_2615081), anti-H3K4me1 (Active Motif Cat# 39297; RRID:AB_2615075), anti-Aebp2 (Cell Signaling Technology Cat# 14129; RRID: AB_2798398), anti-H3K27me3 (Active Motif Cat# 39155; RRID: AB_2561020), anti-H3 (Active Motif Cat# 39763; RRID:AB_2650522), and anti-Jarid2 (Novus Cat# NB100-2214; RRID:AB10000529).
Hickey G.J., Wike C.L., Nie X., Guo Y., Tan M., Murphy P.J, & Cairns B.R. (2022). Establishment of developmental gene silencing by ordered polycomb complex recruitment in early zebrafish embryos. eLife, 11, e67738.
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