• USA Home
  • 17-622 - ChIPAb+ Trimethyl-Histone H3 (Lys27) - ChIP Validated Antibody and Primer Set

EMAIL THIS PAGE TO A FRIEND
17-622 Sigma-Aldrich

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

Synonym: H3K27me3, Histone H3 (tri methyl K27), Histone H3K27me3

  •  eCl@ss 32160702

  •  NACRES NA.32

Purchase

Properties

Related Categories Antibodies, Antibodies for Epigenetics and Nuclear Signaling, ChIP Validated Antibodies, Primary Antibodies
clone   polyclonal
biological source   rabbit
application(s)   ChIP: suitable
  western blot: suitable
species reactivity   human, mouse
shipped in   dry ice
Quality Level   100
purified by   using Protein A
mfr. no.   ChIPAb+
  Upstate®
NCBI accession no.   NP_003484
UniProt accession no.   Q16695
Gene Information   human ... H3F3B(3021)

Description

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+ Trimethyl-Histone H3 (Lys27) set includes the Anti-Trimethyl-Histone H3 (Lys27) antibody, a negative control antibody (purified rabbit IgG), and qPCR primers which amplify a 139 bp region of human alpha-Satellite. The Trimethyl-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 Trimethyl-Histone H3 (Lys27)-associated chromatin.

The methylation of histones can occur on two different residues: arginine or lysine. Histone methylation can be associated with transcriptional activation or repression, depending on the methylated residue. Lysine 27 of histone H3 can be mono-, di- or trimethylated (Histone H3 monomethyl Lys27, Histone H3 dimethyl Lys27 or Histone H3 trimethyl Lys27) by different histone methyltransferases such as EZH2 or NSD3. Methylation of this residue is mainly associated with transcriptional repression.

Specificity

Broad species cross-reactivity expected.

trimethyl-Histone H3 (Lys27)

Immunogen

Epitope: Trimethyl Lys27

KLH-conjugated, synthetic 2X-branched peptide containing the sequence ...AR(me3K)SAP... in which me3K corresponds to trimethyl-lysine at residue 27 of human Histone H3.

Application

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from 2x106 HeLa cells were subjected to chromatin immunoprecipitation using 4 μg purified anti-trimethyl-Histone H3 (Lys27) antibody or normal rabbit IgG and the Magna ChIP A kit (Cat. #17-610). Successful enrichment of trimethyl-Histone H3 (Lys27) associated DNA fragments was verified by qPCR using ChIP Primers GAPDH flanking the human GAPDH promoter and primers targeting the promoter of human MyoD.
Please refer to the EZ-Magna ChIP A (Cat. #17-408) or EZChIP (Cat. #17-371) kit protocols for experimental details.

Western Blot Analysis:
Recombinant Histone H3 (Lane 2) and HeLa cell acid extracts (Lane 1) were resolved by electrophoresis, transferred to nitrocellulose and probed with a 1:5000 dilution of anti- trimethyl histone H3 Lys27 (0.4 μg/mL).
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
Histones

Trimethyl-Histone H3 (Lys27) ChIP validated antibody & primer set including the ChIP-grade antibody & the specific control PCR primers used for chromatin immunoprecipitation of H3K27Me3.

Target description

17kDa

Physical form

Anti-trimethyl-Histone H3 (Lys27) (rabbit polyclonal IgG). One vial containing 100 μg protein A purified IgG in 100 μL buffer containing 0.1 M Tris-Glycine, pH 7.4, 0.15M NaCl with 0.05% sodium azide. Store at -20°C.

Normal Rabbit IgG. One vial containing 125 μg purified Rabbit IgG in 125 μL storage buffer containing 0.1% sodium azide. Store at -20°C.

ChIP Primers human alpha-Satellite. One vial containing 75 μL of 5 μM of each control primer specific for human alpha-Satellite. Store at -20°C.
FOR: CTG CAC TAC CTG AAG AGG AC
REV: GAT GGT TCA ACA CTC TTA CA

Storage and Stability

1 year at -20°C from date of shipment

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.

Legal Information

MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany

Packaging

25 assays per kit, ~4μL per chromatin immunoprecipitation.

Components

Anti-trimethyl Histone H3(Lys27) polyclonal, 1 vial

Polyclonal control rabbit serum, 1 vial

Human alpha-Satellite primer set, 1 vial

Quality

Chromatin Immunoprecipitation:
Sonicated Chromatin prepared from 3x106 NIH3T3 L1 cells were subjected to chromatin immunoprecipitation using 4 μg purified antibody or normal rabbit IgG and the Magna ChIP® A kit (Cat. #17-610). Successful enrichment of trimethyl-Histone H3
(Lys27) associated DNA fragments was verified by qPCR using ChIP Primers human alpha-Satellite (Please see figures).
Please refer to the EZ-Magna ChIP A (Cat. #17-408) or EZ-ChIP (Cat. #17-371) kit protocols for experimental details.

Analysis Note

Control
1 vial containing 75μl of 5μM of each control primer specific for human alpha-Satellite. Store at -20°C.

FOR: CTG CAC TAC CTG AAG AGG AC

REV: GAT GGT TCA ACA CTC TTA CA

Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.

Documents

Certificate of Analysis (COA)

Please Enter a Lot Number
Protocols & Articles

Articles

Antibody Basics

Immunoglobulins (Igs) are produced by B lymphocytes and secreted into plasma. The Ig molecule in monomeric form is a glycoprotein with a molecular weight of approximately 150 kDa that is shaped more ...
Keywords: Affinity chromatography, Centrifugation, Chromatography, Digestions, Direct immunofluorescence, Gene expression, High performance liquid chromatography, Immunofluorescence, Ion Exchange, Microscopy, Precipitation, Purification, Rheumatology, Scanning electron microscopy

Protocols

Western Blot Protocol | Immunoblotting Protocol

Western Blotting refers to the electrophoretic transfer of proteins from sodium dodecyl sulfate polyacrylamide gels to sheets of PVDF or nitrocellullose membrane, followed by immunodetection of prote...
Keywords: AGE, Buffers, Cell disruption, Detection methods, Detergents, Dialysis, Electroblotting, Electrophoresis, Enzyme activity, Gel electrophoresis, Immunoprecipitation, PAGE, Protein extraction, Purification, Sample preparations, Western blot

Related Content

Antibody Explorer | Buy Primary Antibodies & Secondary Antibodies

Primary, Secondary and Recombinant Monoclonal Antibodies Providing highly cited primary and secondary antibodies, we have you covered for your ELISA, western blot, immunohistochemistry or other assay...
Keywords: Alzheimer Disease, Cancer, Chromatin immunoprecipitation, Diagnostic, Enzyme-linked immunosorbent assay, Flow cytometry, Immunocytochemistry, Immunofluorescence, Immunohistochemistry, Neuroscience, Western blot

Peer-Reviewed Papers
15

References

Related Products

Technical Service:

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Bulk Ordering & Pricing:

Need larger quantities for your development, manufacturing or research applications?