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ChIPAb+ Dimethyl-Histone H3 (Lys9) - ChIP Validated Antibody and Primer Set
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REFERENCES
The organization of histone H3 modifications as revealed by a panel of specific monoclonal antibodies.
Kimura, Hiroshi, et al. (2008) Cell Struct. Funct., 33: 61-73 (2008)
Species Reactivity Key Applications Host Format Antibody Type
Vrt WB, ChIP Mouse null Monoclonal Antibody
Description:
ChIPAb+ Dimethyl-Histone H3 (Lys9)
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Trade Name:
Upstate (Millipore)
Product Overview:
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+ Dimethyl-Histone H3 (Lys9) set includes the Anti-dimethyl-Histone H3 (Lys9) antibody, a negative control antibody (purified Mouse IgG), and qPCR primers which amplify a 110 bp region within the promoter of the human β-globin gene. The dimethyl-histone H3 (Lys9) and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of dimethyl-histone H3 (Lys9) associated chromatin.
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Specificity:
Recognizes histone H3, Mr 17 kDa, dimethylated at lysine 9.
Molecular Weight:
Dimethyl-histone H3 at ~17 kDa
Epitope:
a.a. 1-18
Immunogen:
The dimethyl-histone H3 (Lys9) purified antibody is made against a synthetic peptide (dimethylated at Lys9) corresponding to amino acids 1-18 of histone H3.
Modifications:
Methylation
Isotype:
IgG
Species Reactivity:
Vertebrates
Species Reactivity Note:
Human, although this peptide sequence is identical in a wide range of animal and plant species.
Application Notes:
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-dimethyl-Histone H3 (Lys9) antibody and the Magna ChIP G (Cat. #17-611) Kit. Successful immunoprecipitation of dimethyl-histone H3 (Lys9) associated DNA fragments was verified by qPCR using β-globin ChIP Primers versus Control 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:
Recombinant Histone H3 (Lane 1) and HeLa acid extract (Lane 2) were resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-dimethyl Histone H3 (Lys9), clone CMA307 (0.1 μg/ml). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system.
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Control:
Included negative control mouse IgG antibody and control primers specific for human β-globin promoter.
Quality Assurance:
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-dimethyl-Histone H3 (Lys9) antibody and the Magna ChIP G (Cat. #17-611) Kit. Successful immunoprecipitation of dimethyl-histone H3 (Lys9) associated DN |