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17-643 ChIPAb+ Monomethyl-Histone H3 (Lys27) - ChIP Validated Antibody and Primer Set

This ChIPAb+ Monomethyl-Histone H3 (Lys27) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Key Applications: Flow Cytometry, Electrophoretic Mobility Shift Assay, Immunofluorescence, Immunohistochemistry, Immunohistochemistry (Paraffin), Western Blotting, Chromatin Immunoprecipitation (ChIP) Research Category: Epigenetics & Nuclear Function Research Sub-Category: Chromatin Biology
17-643
25 assays  25 assays per kit, ~4μL per chromatin immunoprecipitation
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Key Spec Table

Species ReactivityKey Applications
H, M, MaFC, EMSA, IF, IHC, IH(P), WB, ChIP
Description
Catalogue Number17-643
Brand Family Upstate
Trade Name
  • ChIPAb+
  • Upstate
DescriptionChIPAb+ Monomethyl-Histone H3 (Lys27) - ChIP Validated Antibody and Primer Set
OverviewAll 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.
Alternate Names
  • H3K27me1
  • Histone H3 (mono methyl K27)
Product Information
FormatSerum
Control
  • Included negative control antibody normal rabbit serum and control primers specific for human β-globin promoter.
PresentationAnti-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
Quality LevelMQ100
Applications
ApplicationThis ChIPAb+ Monomethyl-Histone H3 (Lys27) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Key Applications
  • Flow Cytometry
  • Electrophoretic Mobility Shift Assay
  • Immunofluorescence
  • Immunohistochemistry
  • Immunohistochemistry (Paraffin)
  • Western Blotting
  • Chromatin Immunoprecipitation (ChIP)
Application NotesChromatin 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).
Biological Information
ImmunogenThe 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.
HostRabbit
SpecificityMonomethyl-Histone H3 (Lys27)
Species Reactivity
  • Human
  • Mouse
  • Mammals
Antibody TypePolyclonal Antibody
Entrez Gene Number
Gene Symbol
  • H3F3A
  • H3F3B
  • H3F3
UniProt Number
UniProt SummaryFunction: Variant histone H3 which replaces conventional H3 in a wide range of nucleosomes in active genes. Constitutes the predominant form of histone H3 in non-dividing cells and is incorporated into chromatin independently of DNA synthesis. Deposited at sites of nucleosomal displacement throughout transcribed genes, suggesting that it represents an epigenetic imprint of transcriptionally active chromatin. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.
Subunit structure: The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA. Interacts with HIRA, a chaperone required for its incorporation into nucleosomes.
Subcellular location: Nucleus.
Developmental stage: Expressed throughout the cell cycle independently of DNA synthesis.
Post-translational modification: Acetylation is generally linked to gene activation. Acetylation on Lys-10 (H3K9ac) impairs methylation at Arg-9 (H3R8sme2). Acetylation on Lys-19 (H3K18ac) and Lys-24 (H3K24ac) favors methylation at Arg-18 (H3R17me).
Citrullination at Arg-9 (H3R8ci) and/or Arg-18 (H3R17ci) by PADI4 impairs methylation and represses transcription.
Asymmetric dimethylation at Arg-18 (H3R17me2a) by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 (H3R8sme2) by PRMT5 is linked to gene repression. Asymmetric dimethylation at Arg-3 (H3R2me2a) by PRMT6 is linked to gene repression and is mutually exclusive with H3 Lys-5 methylation (H3K4me2 and H3K4me3). H3R2me2a is present at the 3' of genes regardless of their transcription state and is enriched on inactive promoters, while it is absent on active promoters.
Specifically enriched in modifications associated with active chromatin such as methylation at Lys-5 (H3K4me), Lys-37 and Lys-80. Methylation at Lys-5 (H3K4me) facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 (H3K79me) is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me), which are linked to gene repression, are underrepresented. Methylation at Lys-10 (H3K9me) is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 (H3S10ph) and acetylation of H3 and H4. Methylation at Lys-5 (H3K4me) and Lys-80 (H3K79me) require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are enriched in inactive X chromosome chromatin.
Phosphorylated at Thr-4 (H3T3ph) by GSG2/haspin during prophase and dephosphorylated during anaphase. At centromeres, specifically phosphorylated at Thr-12 (H3T11ph) from prophase to early anaphase, probably DAPK3. Phosphorylation at Ser-11 (H3S10ph) by AURKB is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at Ser-11 (H3S10ph) by RPS6KA4 and RPS6KA5 is important during interphase because it enables the transcription of genes following external stimulation, like mitogens, stress, growth factors or UV irradiation and result in the activation of genes, such as c-fos and c-jun. Phosphorylation at Ser-11 (H3S10ph), which is linked to gene activation, prevents methylation at Lys-10 (H3K9me) but facilitates acetylation of H3 and H4. Phosphorylation at Ser-11 (H3S10ph) by AURKB mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. Phosphorylation at Ser-11 (H3S10ph) is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylated at Ser-29 by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation. Phosphorylation on Ser-32 is specific to regions bordering centromeres in metaphase chromosomes.
Ubiquitinated. Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination.
Sequence similarities: Belongs to the histone H3 family.
Molecular Weight17 kDa
Product Usage Statements
Quality AssuranceChromatin 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).
Usage Statement
  • 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 and Shipping Information
Storage ConditionsStable for 1 year at -20°C from date of receipt
Packaging Information
Material Size25 assays
Material Package25 assays per kit, ~4μL per chromatin immunoprecipitation
Global Trade Item Number
Catalogue Number GTIN
17-643 04053252475108