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Merck

17-683

ChIPAb+ Acetyl-Histone H3 (Lys27) - ChIP Validated Antibody and Primer Set

clone CMA309, from mouse

Synonyme(s) :

H3K27Ac, Histone H3 (acetyl K27), Histone H3K27Ac

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A propos de cet article

UNSPSC Code:
12352203
NACRES:
NA.32
eCl@ss:
32160702
Clone:
CMA309, monoclonal
Species reactivity:
vertebrates, human
Application:
ChIP, IP, WB
Citations:
26

Principaux documents

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biological source

mouse

antibody form

purified immunoglobulin

clone

CMA309, monoclonal

species reactivity

vertebrates, human

manufacturer/tradename

ChIPAb+, Upstate®

technique(s)

ChIP: suitable, immunoprecipitation (IP): suitable, western blot: suitable

isotype

IgG

NCBI accession no.

NM_003493

UniProt accession no.

P84243, Q66I33

shipped in

dry ice

Quality Level

100

General description

All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
The ChIPAb+ Acetyl-Histone H3 (Lys27) set includes the Anti-acetyl-Histone H3 (Lys27) antibody, a negative control antibody (purified Mouse IgG), and qPCR primers which amplify a 178 bp region within the promoter of the human RPL10 gene. The acetyl-histone H3 (Lys27) and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of acetyl-histone H3 (Lys27)-associated chromatin.
Lysine acetylation is a dynamic, reversible and tightly regulated protein and histone modification that plays a major role in chromatin remodeling and in the regulation of gene expression in various cellular functions. Histone acetylation is often associated with transcriptional activation and acetylation of H3K27 is found at active enhancers.
The previously assigned protein identifier Q66I33 has been merged into P84243. Full details can be found on the UniProt database.
~17 kDa

Immunogen

Epitope: a.a. 19-37
The acetyl-histone H3 (Lys27) purified antibody is made against a synthetic peptide (acetylated at Lys27) corresponding to amino acids 19-37 of histone H3.

Application

Acetyl-Histone H3 (Lys27) ChIP validated antibody & primer set including the ChIP-grade antibody & the specific control PCR primers used for chromatin immunoprecipitation of H3K27Ac.
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 2 µg of either a normal mouse IgG or Anti-acetyl-Histone H3 (Lys27) antibody and the Magna ChIP G (Cat. #17-611) Kit. Successful immunoprecipitation of acetyl-histone H3 (Lys27)-associated DNA fragments was verified by qPCR using β-globin Promoter ChIP Primers versus RPL10 Promoter Primers (Please see figures). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
Please refer to the EZ-Magna G ChIP (Cat. #17-408) or EZ-ChIP (Cat. #17-371) protocol for experimental details.

Western Blot Analysis:
Acid extracts from untreated (Lane 1) and sodium-butyrate treated (Lane 2) HeLa cells were resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-acetyl Histone H3 (Lys27), clone CMA309 (0.1 μg/mL). Proteins were visualized using a goat
anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures).
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Chromatin Biology

Biochem/physiol Actions

Recognizes histone H3, Mr 17 kDa, acetylated at Lys27.
This peptide sequence is identical in a wide range of animal and plant species.

Packaging

25 assays per kit, ~2μg per chromatin immunoprecipitation

Physical form

Anti-acetyl-Histone H3 (Lys27) (mouse monoclonal IgG1, Clone CMA309). One vial containing 50 μg of protein G purified antibody in 50 μL PBS containing 0.05% sodium. Store at -20°C.

Normal Mouse IgG. Two vials containing 25 μg purified Mouse IgG in 25 μL storage buffer containing 0.1% sodium azide. Store at -20°C.

ChIP Primers, RPL10 Promoter. One vial containing 75 μL of 5 μM of each primer specific for the promoter region of human RPL10. Store at -20°C.
FOR: ACC CGT CTT CGA CAG GAC T
REV: GGA ACG GAA GAC GAG AAC AG
Format: Purified

Preparation Note

Stable for 1 year at -20°C from date of receipt. Aliquot upon thawing, avoid freeze thaw cycles.

Analysis Note

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either a normal mouse IgG or Anti-acetyl-Histone H3 (Lys27) antibody and the Magna ChIP G (Cat. #17-611) Kit.
Successful immunoprecipitation of acetyl-histone H3 (Lys27) associated DNA fragments was verified by qPCR using ChIP Primers RPL10 Promoter (Please see figures).
Please refer to the EZ-Magna G ChIP (Cat. #17-409) or EZ-ChIP (Cat. #17-371) protocol for experimental details.
Control
Included negative control mouse IgG antibody and control primers specific for human RPL10 promoter.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Classe de stockage

10 - Combustible liquids

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Marta Kubiak et al.
International journal of molecular sciences, 20(2) (2019-01-13)
The long-range control of gene expression is facilitated by chromatin looping and can be detected using chromosome conformation capture-3C. Here we focus on the chromatin architecture of the PTBP3 (Polypyrimidine tract binding protein 3) locus to evaluate its potential role
Katherine A Pillman et al.
Nucleic acids research, 47(16), 8606-8619 (2019-08-03)
Epithelial-mesenchymal transition (EMT) has been a subject of intense scrutiny as it facilitates metastasis and alters drug sensitivity. Although EMT-regulatory roles for numerous miRNAs and transcription factors are known, their functions can be difficult to disentangle, in part due to
Akua Yalley et al.
The Journal of biological chemistry, 291(16), 8848-8861 (2016-03-02)
FoxO1 binds to insulin response elements located in the promoters of insulin-like growth factor-binding protein 1 (IGFBP1) and glucose-6-phosphatase (G6Pase), activating their expression. Insulin-mediated phosphorylation of FoxO1 promotes cytoplasmic translocation, inhibiting FoxO1-mediated transactivation. We have previously demonstrated that FoxO1 opens
Chao Yang et al.
Molecular plant, 13(3), 515-531 (2020-02-23)
Light is arguably one of the most important environmental factors that determines virtually all aspects of plant growth and development, but the molecular link between light signaling and the autophagy pathway has not been elucidated in plants. In this study
Joseph Cursons et al.
Cell systems, 7(1), 77-91 (2018-07-17)
MicroRNAs (miRNAs) are important post-transcriptional regulators of gene expression, functioning in part by facilitating the degradation of target mRNAs. They have an established role in controlling epithelial-mesenchymal transition (EMT), a reversible phenotypic program underlying normal and pathological processes. Many studies
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Advance your Epigenetics Research

Epigenetics describes heritable changes in gene expression caused by non-genetic mechanisms. Epigenetic regulation allows a cell to vary its response based on its biological and environmental contexts. Epigenetic changes can effect transcriptional and post-transcriptional regulation via mechanisms such as histone modification, chromatin and nucleosome remodeling, DNA methylation, and small and non-coding RNA-mediated regulation. These mechanisms, in cooperation with transcription factors and other nucleic acid-binding proteins, regulate gene expression. Epigenetic mechanisms of gene regulation impacts diverse areas of research—from agriculture to human health. Common epigenetic assays such as chromatin immunoprecipitation (ChIP) and RNA immunoprecipitation (RIP) rely on high quality antibodies that recognize specific epigenetic modifications for accurate results. EMD Millipore offers over 100 ChIPAb+™ and RIPAb+™ validated antibody kits that are quality tested on ChIP/RIP assays and are conveniently provided with control qPCR primers and negative control antibodies to ensure first time ChIP/RIP success.

Shaping Epigenetics Discovery - Epigenetics Product Selection Brochure

"Epigenetics describes heritable changes in gene expression caused by non-genetic mechanisms instead of by alterations in DNA sequence. These changes can be cell- or tissue-specific, and can be passed on to multiple generations. Epigenetic regulation enriches DNAbased information, allowing a cell to vary its response across diverse biological and environmental contexts. Although epigenetic mechanisms are primarily centered in the nucleus, these mechanisms can be induced by environmental signals such as hormones, nutrients, stress, and cellular damage, pointing to the involvement of cytoplasmic and extracellular factors in epigenetic regulation."

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