H3k4me1 ab8895

Manufactured by Abcam

Sourced in Morocco, United Kingdom

H3K4me1 (ab8895) is a primary antibody that specifically recognizes the monomethylation of lysine 4 on histone H3. This epigenetic modification is associated with active gene transcription.

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Anti-H3K4me1 (ab8895) Ab8895 H3K4me1

5 protocols using h3k4me1 ab8895

Quantitative Immunohistochemical Analysis of Tongue Epithelium

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Experiments were performed as previously described (Osei-Sarfo et al., 2013 (link), Tang et al., 2014 ). Sections were incubated with primary antibodies overnight at 4 °C (1:200 TOM20 (sc-11415, Santa Cruz, Santa Cruz, CA), 1:200 4-HNE (Ab48506, Abcam, Cambridge, MA), 1:100 MCT4 (sc-50329, Santa Cruz), 1:400 H3K27ac (Ab4729, Abcam), 1:1,000 H3K9/14ac (06-599, Millipore, Billerica, MA), 1:400 H3K27me3 (07-449, Millipore), 1:1,000 H3K9me3 (07-473, Millipore), 1:1,000 H3K4me1 (Ab8895, Abcam)). The antibody signal was visualized by peroxidase reaction using 3,3′-diaminobenzidine. Methyl green was used as a nuclear counterstain. At least three mice were used for each treatment group, and for each sample at least four non-contiguous regions were photographed and analyzed. Image J (http://imagej.nih.gov) software was used to measure positively stained tongue epithelial area compared to total epithelial area. Ratios were calculated and converted to percent positive area.

Urvalek A.M., Osei-Sarfo K., Tang X.H., Zhang T., Scognamiglio T, & Gudas L.J. (2015). Identification of Ethanol and 4-Nitroquinoline-1-Oxide Induced Epigenetic and Oxidative Stress Markers During Oral Cavity Carcinogenesis. Alcoholism, clinical and experimental research, 39(8), 1360-1372.

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Chromatin Modification Profiling

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ERα ab32063 ((Abcam, Cambridge, UK); ERβ ab288 (Abcam); H3K4me1 ab8895 (Abcam); H3K4me2 ab32356 (Abcam); H3K4me3 ab1012 (Abcam); H3K9me2 ab1220 (Abcam); H3K9me3 ab8898 (Abcam); H3K27Ac ab4729 (Abcam); Total H3 ab1791 (Abcam); LSD1 ab17721 (Abcam); JMJD2A sc-271,210 (Santa Cruz Biotechnology); Normal rabbit IgG sc-2027 (Santa Cruz Biotechnology); Normal mouse IgG sc-2025 (Santa Cruz Biotechnology).

Romano A., Feola A., Porcellini A., Gigantino V., Di Bonito M., Di Mauro A., Caggiano R., Faraonio R, & Zuchegna C. (2020). Estrogen Induces Selective Transcription of Caveolin1 Variants in Human Breast Cancer through Estrogen Responsive Element-Dependent Mechanisms. International Journal of Molecular Sciences, 21(17), 5989.

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Chromatin Immunoprecipitation Techniques for Epigenetic Regulation

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ChIP, ChIP-Seq and sequential-ChIP assays were performed as described previously (6 (link),14 (link)). Briefly, Nuclei were sonicated with the Bioruprtor™ UCD200. Chromatin samples prepared from 1 × 10⁶ cells of Day 19 EBs or bone marrow (BM) cells were immunoprecipitated with antibodies specific for histone modifications or epigenetic regulators. The immunoprecipitated chromatin complexes were reverse cross-linked and purified. The recovered DNA was analyzed by qPCR. The relative enrichment was determined by the following equation: 2^{Ct(IP)-Ct(ref)}. The sequential ChIP assays were carried out essentially as described (14 (link),27 (link)) with some modifications. Briefly, chromatin prepared from 5 × 10⁶ cells was immunoprecipitated with antibodies specific for H3K4me3 or USF1. The H3K4me3 or USF1-selected chromatin complexes were eluted, dialyzed and subsequently immunoprecipitated with antibody specific to BPTF. The bound protein–DNA complexes were reverse cross-linked and purified. The recovered DNA was analyzed by qPCR.
Antibodies against H3K4me2 (07-730), H3K4me3 (04-745), SNF2L (05-698) and BPTF (ABE24) were purchased from Millipore, H3K4me1 (ab8895) was from Abcam, ASH2L (A300-489A) and Setd1a (A300-289A) were from Bethyl laboratories and USF1 (H-86) was from Santa Cruz Biotechnology.

Li Y., Schulz V.P., Deng C., Li G., Shen Y., Tusi B.K., Ma G., Stees J., Qiu Y., Steiner L.A., Zhou L., Zhao K., Bungert J., Gallagher P.G, & Huang S. (2016). Setd1a and NURF mediate chromatin dynamics and gene regulation during erythroid lineage commitment and differentiation. Nucleic Acids Research, 44(15), 7173-7188.

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MNase-ChIP-seq and MNase-seq Protocols

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MNase-ChIP-seq and MNase-seq protocols were performed according to previous studies^{44 (link)}. In brief, LNCaP cells were exposed to 10 nM DHT or DMSO (Veh) for 4 h. Mono-nucleosomes with solubilized chromatin was achieved by MNase digestion of 2 min at 37 °C, then immunoprecipitated with antibody-conjugated magnetic beads. DNA is phenol extracted and ethanol precipitated. Libraries were prepared from isolated DNA and sent for sequencing on the Illumina HiSeq3000 at the UTHSA sequencing core. All samples were performed in biological replicates. Antibodies include: H3K4me1 (ab8895) 1:500 dilution, H3K4me2 (ab7766) 1:250 dilution, H3K27ac (ab4729) 1:500 dilution, H3K27me3 (ab6002) 1:250 dilution, H3K36me3 (ab9050) 1:250 dilution, H3K79me2 (ab8898) 1:250 dilution from Abcam (Cambridge, MA). H3K4me3 (07-473) 1:500 dilution, H3K9me3 (17-10242) 1:250 dilution from Millipore (Upstate).

Li T., Liu Q., Chen Z., Fang K., Huang F., Fu X., Wang Q, & Jin V.X. (2022). Dynamic nucleosome landscape elicits a noncanonical GATA2 pioneer model. Nature Communications, 13, 3145.

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Western Blot and ChIP Antibody Panel

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The following antibodies were used in western blot and ChIP: TRIM33 (3E9, Euromedex for WB (1:1,000); A301-059A, Bethyl for ChIP (1:100)), c-JUN (sc-1694X, Santa Cruz (1:200)), p65 (sc-372X, Santa Cruz (1:200)), RNA Pol II (sc-899X, Santa Cruz (1:200)), H3K4me3 (07-473, Millipore (1:500)), PU.1 (sc-352X, Santa Cruz (1:10,000 for western blot; 1:500 for ChIP)), p300 (sc-585X, Santa Cruz), AcH3 (06-599, Millipore (1:500)), H3K4me1 (ab-8895, Abcam (1:500)), Flag (M2 monoclonal, Sigma Aldrich (1:1,000)), CBP (A-22, sc-369, Santa Cruz (1:1,000)), β-actin (A5441, Sigma Aldrich (1:10,000)).

Ferri F., Parcelier A., Petit V., Gallouet A.S., Lewandowski D., Dalloz M., van den Heuvel A., Kolovos P., Soler E., Squadrito M.L., De Palma M., Davidson I., Rousselet G, & Romeo P.H. (2015). TRIM33 switches off Ifnb1 gene transcription during the late phase of macrophage activation. Nature Communications, 6, 8900.

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