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Anti h3k4me1 ab8895

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Anti-H3K4me1 (ab8895) is a primary antibody that recognizes the monomethylation of lysine 4 on histone H3. It is designed for use in various applications such as Western blotting, immunoprecipitation, and chromatin immunoprecipitation.

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Lab products found in correlation

Formaldehyde, by Thermo Fisher Scientific (1 mentions) Anti gr, by Cell Signaling Technology (1 mentions) Zymo spin chip kit, by Zymo Research (1 mentions) Anti h3k4me3, by Cell Signaling Technology (1 mentions) Anti h3k27me3, by Cell Signaling Technology (1 mentions) Anti h3k27ac, by Cell Signaling Technology (1 mentions) Anti h3k27me3 c15410195, by Diagenode (1 mentions) Anti grhl2 hpa004820, by Merck Group (1 mentions) Anti h3k4me3 ab8580, by Abcam (1 mentions) Anti h3k4me2 07 030, by Merck Group (1 mentions) Anti histone h3 ab1791, by Abcam (1 mentions) Anti h3k27ac ab4729, by Abcam (1 mentions)

Spelling variants of this product

Ab8895 (162 citations) H3K4me1 (98 citations) Anti-H3K4me1 (42 citations) H3K4me1 (ab8895) (5 citations)

5 protocols using anti h3k4me1 ab8895

1

Chromatin Immunoprecipitation Assay Protocol

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ChIP assay was performed using the Zymo-Spin ChIP kit according to the manufacturer’s protocol (Zymo Research). Briefly, FLS were serum starved for 24 hours and then fixed in 1% formaldehyde (Thermo Fisher Scientific) for 8 minutes at room temperature. After sonication, chromatin was immunoprecipitated with specific antibodies overnight at 4°C. The antibodies used were anti-H3K4me1 (ab8895, Abcam), anti-GR (catalog 12041, Cell Signaling Technology), and rabbit IgG (catalog 2729, Cell Signaling Technology). Complexes were then immunoprecipitated with protein A magnetic beads for 1 hour at 4°C and reverse-crosslinked at 65°C for 2 hours. The eluted DNA was purified and used as template for qPCR. A 10% input sample was used as control. The enhancer-specific primers, 5′-CTGAGACTGGAGGCTTTTGG-3′ (forward) and 5′-GACATCCCAGTTTCCACTGC-3′ (reverse), were designed using Primer3 software (42 (link)). MYT1 exon 1 primers were purchased from Cell Signaling Technology (catalog 4493).
Maeshima K., Stanford S.M., Hammaker D., Sacchetti C., Zeng L.F., Ai R., Zhang V., Boyle D.L., Aleman Muench G.R., Feng G.S., Whitaker J.W., Zhang Z.Y., Wang W., Bottini N, & Firestein G.S. (2016). Abnormal PTPN11 enhancer methylation promotes rheumatoid arthritis fibroblast-like synoviocyte aggressiveness and joint inflammation. JCI Insight, 1(7), e86580.
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2

Antibody-based ChIP Profiling

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Antibodies used for ChIP consisted of anti-H3K27ac (8173; Cell Signaling Technology, Danvers, MA), anti-H3K4me1 (ab8895; Abcam, Cambridge, MA), anti-H3K4me3 (purified antibody generated in-house by the Mayo Clinic Epigenomics Development Lab, Rochester, MN [60 (link)], and anti-H3K27me3 (9733, Cell Signaling Technology).
Ghaffari S., Hanson C., Schmidt R.E., Bouchonville K.J., Offer S.M, & Sinha S. (2021). An integrated multi-omics approach to identify regulatory mechanisms in cancer metastatic processes. Genome Biology, 22, 19.
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3

ChIP-seq Antibody Characterization

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The antibodies used for ChIP-seq were anti-FOXA1 (ab5089) from Abcam, anti-H3K4me1 (ab8895) from Abcam, anti-H3K4me3 (05-1339) from Millipore, anti-H3K27ac (C15410196) from Diagenode, anti-H3K27me3 (C15410195) from Diagenode, anti-GRHL2 (HPA004820) from Sigma-Aldrich, and negative control immunoglobulin G (IgG) anti-rabbit IgG (sc-2027), and anti-goat (sc-2028) from Santa Cruz Biotechnology. The custom anti-MLL3 antibody was provided by Prof. Ali Shilatifard (Stowers Institute, Kansas City, MO, and Northwestern University Feinberg School of Medicine, Chicago, IL).
Jozwik K.M., Chernukhin I., Serandour A.A., Nagarajan S, & Carroll J.S. (2016). FOXA1 Directs H3K4 Monomethylation at Enhancers via Recruitment of the Methyltransferase MLL3. Cell Reports, 17(10), 2715-2723.
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4

Jarid1B Protein Interaction Analysis

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Cell lysates were prepared 5 days after siRNA transfection. The proteins resolved in SDS – polyacrylamide gels (4–12%) were transferred electrophoretically for 2 hr at 4 °C to polyvinylidene difluoride membranes by using a Tris-glycine buffer system. The membranes were blocked with 5% milk powder in 0.1% Tween20 in PBS (PBS-T) for 1 hr at least, and then the antibodies were added with 2.5% milk in PBS-T. The antibodies used for immunoblotting were anti-Jarid1B (HPA027179) from Sigma, anti-Jarid1A (ab78322), anti-H3K4me3 (ab8580), anti-H3K4me1 (ab8895), anti-Histone H3 (ab1791) from Abcam, anti-H3K4me2 (07-030) from Millipore, and anti-CTCF (BD612148) from BD Biosciences. For co-IP, nuclear extract of cell-lines were prepared as for the ChIP experiments, sheared by passing through syringe needle, diluted, and treated with DNase I. The samples were incubated at 4°C overnight with the JARID1B antibody, and then precipitated with Dynabeads Protein G for 2 hr. Beads were washed with buffer containing 150 mM NaCl and 0.5% NP-40 three times, and then resuspended in SDS – polyacrylamide gel loading buffer.
Yamamoto S., Wu Z., Russnes H.G., Takagi S., Peluffo G., Vaske C., Zhao X., Vollan H.K., Maruyama R., Ekram M.B., Sun H., Kim J.H., Carver K., Zucca M., Feng J., Almendro V., Bessarabova M., Rueda O.M., Nikolsky Y., Caldas C., Liu X.S, & Polyak K. (2014). JARID1B is a luminal lineage-driving oncogene in breast cancer. Cancer cell, 25(6), 762-777.
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5

ChIP-qPCR for H3K4me1 and H3K27Ac

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Chromatin immunoprecipitation experiments were performed exactly as previously described (62 (link)). Briefly, nuclear extracts from 2 × 106 neutrophils or Th17 cell lines were immunoprecipitated using 1 µl anti-H3K4me1 (ab8895) and anti-H3K27Ac (ab4729) pAbs (both from Abcam, Cambridge, United Kingdom). Coimmunoprecipitated material was subjected to qPCR analysis using the specific promoter primers (purchased from Life Technologies) listed in Table S2 in Supplementary Material. Data from qPCR were expressed as percentage over input DNA and are displayed as mean ± SEM.
Tamassia N., Arruda-Silva F., Calzetti F., Lonardi S., Gasperini S., Gardiman E., Bianchetto-Aguilera F., Gatta L.B., Girolomoni G., Mantovani A., Vermi W, & Cassatella M.A. (2018). A Reappraisal on the Potential Ability of Human Neutrophils to Express and Produce IL-17 Family Members In Vitro: Failure to Reproducibly Detect It. Frontiers in Immunology, 9, 795.
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