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Merck

17-643

Sigma-Aldrich

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

serum, from rabbit

Synonym(s):

H3K27me1, Histone H3 (mono methyl K27)

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.52

biological source

rabbit

Quality Level

100

antibody form

serum

clone

polyclonal

species reactivity

human, mouse

species reactivity (predicted by homology)

mammals

manufacturer/tradename

ChIPAb+
Upstate®

technique(s)

ChIP: suitable
electrophoretic mobility shift assay: suitable
flow cytometry: suitable
immunofluorescence: suitable
immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable
immunoprecipitation (IP): suitable
western blot: suitable

NCBI accession no.

NM_003493

UniProt accession no.

Q66I33

shipped in

dry ice

Gene Information

human ... H3F3B(3021)

Related Categories

General description

17 kDa
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+ Monomethyl-Histone H3 (Lys27) set includes the anti-monomethyl-histone H3 (Lys27) antibody, a negative control antibody (normal rabbit serum), and qPCR primers which amplify a 110 base pair region within the promoter of the human β-globin gene. The monomethyl-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 monomethyl-histone H3 (Lys27)-associated chromatin.

Immunogen

The monomethyl-histone H3 (Lys27) antiserum is made against a synthetic 2X-branched peptide containing the sequence…ARmeKSA…in which meK corresponds to monomethyl-lysine 27 of human histone H3.

Application

Chromatin Immunoprecipitation:
Successful immunoprecipitation of monomethyl-histone H3 (Lys27) associated DNA fragments was verified by qPCR using control ChIP Primers flanking the human GAPDH promoter or primers amplifying the promoter of human β-globin, which is transcriptionally inactive in HeLa cells (Please see figures). Percent Input relative to standard curves for each qPCR primer set are shown, with immunoprecipitated DNA from control serum shown as (-) and monomethyl- histone H3 (Lys27) as (+).
Please refer to the EZ-Magna A ChIP (Cat. #17-408) or EZ-ChIP (Cat. #17-371) protocol for experimental details.

Western blot analysis and peptide inhibition:
HeLa Acid extract was resolved by electrophoresis, transferred to nitrocellulose and probed with Anti-Monomethyl-Histone H3 (Lys27) (1:5000, Lane 1) or preincubated with 0.4 μM Histone H3 peptide with following modifications:
Lane 2: Linear non-modified
Lane 3: Branched monomethyl
Lane 4: Linear monomethyl
Lane 5: Branched dimethyl
Lane 6: Linear dimethyl
Lane 7: Branched trimethyl
Lane 8: Linear trimethyl
Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures).
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Chromatin Biology
This ChIPAb+ Monomethyl-Histone H3 (Lys27) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Biochem/physiol Actions

Monomethyl-Histone H3 (Lys27)

Packaging

25 assays per kit, ~4μL per chromatin immunoprecipitation

Physical form

Anti-monomethyl-Histone H3 (Lys27) (rabbit polyclonal IgG). One vial containing 100 μL of antiserum containing 0.05% sodium azide.

Normal Rabbit Serum. One vial containing 100 uL antiserum containing 0.05% sodium azide.

ChIP Primers, β-Globin. One vial containing 75 μL of 5 μM of each primer specific for for human β-globin promoter.
FOR: AGG ACA GGT ACG GCT GTC ATC
REV: TTT ATG CCC AGC CCT GGC TC

Preparation Note

Stable for 1 year at -20°C from date of receipt

Analysis Note

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from untreated HeLa cells (1 X 106 cell equivalents) was subjected to chromatin immunoprecipitation using 4 μL of either a normal rabbit antiserum or Anti-Monomethyl-Histone H3 (Lys27) serum and the Magna ChIP A Kit (Cat. #17-610).
Successful immunoprecipitation of monomethyl-histone H3 (Lys27) associated DNA fragments was verified by qPCR using control ChIP Primers flanking the human β-globin promoter (Please see figures).
Please refer to the EZ-Magna A ChIP (Cat. #17-408) or EZ-ChIP (Cat. #17-371).
Control
Included negative control antibody normal rabbit serum and control primers specific for human β-globin 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.

Storage Class Code

10 - Combustible liquids

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Pierre Bourguet et al.
Nature communications, 12(1), 2683-2683 (2021-05-13)
In flowering plants, heterochromatin is demarcated by the histone variant H2A.W, elevated levels of the linker histone H1, and specific epigenetic modifications, such as high levels of DNA methylation at both CG and non-CG sites. How H2A.W regulates heterochromatin organization
An Yan et al.
Development (Cambridge, England), 147(11) (2020-05-23)
Plants are capable of regenerating new organs after mechanical injury. The regeneration process involves genome-wide reprogramming of transcription, which usually requires dynamic changes in the chromatin landscape. We show that the histone 3 variant HISTONE THREE RELATED 15 (H3.15) plays
Zdravko J Lorković
Plant signaling & behavior, 7(12), 1561-1565 (2012-10-18)
Two recent studies in Arabidopsis implicated MORC proteins, which contain a GHKL ATPase domain, in transcriptional gene silencing. Here, these studies are compared and contrasted to discuss the roles of MORC proteins in epigenetic regulation. Although MORC proteins are likely
Bhagyshree Jamge et al.
eLife, 12 (2023-07-19)
How different intrinsic sequence variations and regulatory modifications of histones combine in nucleosomes remain unclear. To test the importance of histone variants in the organization of chromatin we investigated how histone variants and histone modifications assemble in the Arabidopsis thaliana
Michael Borg et al.
Nature cell biology, 22(6), 621-629 (2020-05-13)
Epigenetic marks are reprogrammed in the gametes to reset genomic potential in the next generation. In mammals, paternal chromatin is extensively reprogrammed through the global erasure of DNA methylation and the exchange of histones with protamines1,2. Precisely how the paternal
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Global Trade Item Number

SKUGTIN
17-64304053252475108

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