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ChIPAb+ Histone H3 (Unmodified Lys4) - 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 Protein G Purified Monoclonal Antibody
Description:
ChIPAb+ Histone H3 (Unmodified Lys4) - ChIP Validated Antibody and Primer Set
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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+ Histone H3 (Unmodified Lys4) set includes the Anti-Histone H3 (unmodified Lys4) 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 histone H3 (Unmodified Lys4) and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of histone H3 (Unmodified Lys4)-associated chromatin.
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Specificity:
Recognizes unmodified histone H3 (Lys4), Mr 17 kDa. Epitope specificity of antibody as determined by ELISA maps near Lys4. Antibody tolerates modifications on Lys9 and Ser10 but not Lys4 or Thr31.
Molecular Weight:
Histone H3 at ~17 kDa
Epitope:
a.a. 1-12
Immunogen:
The histone H3 (Unmodified Lys4) purified antibody is made against a synthetic peptide corresponding to amino acids 1-12 of histone H3.
Isotype:
IgG2bκ
Species Reactivity:
Vertebrates
Species Reactivity Note:
Human. The peptide sequence is identical in a wide range of animal and plant species, so broad cross-reactivity is expected.
Application Notes:
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (2 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 2 µg of either normal mouse IgG or Anti- Histone H3 (Unmodified Lys4) antibody and the Magna ChIP G Kit (Cat. #17-611). Successful immunoprecipitation of histone H3 associated DNA fragments was verified by qPCR using β-globin ChIP Primers versus GAPDH Promoter Control Primers. Data is presented as percent input of each IP sample relative to input chromatin, with the indicated primer sets (Please see figures).
Western Blot Analysis:
Acid extracted proteins from HeLa cells (Lane 1) and Recombinant Histones (Lane 2-6) were resolved by electrophoresis, transferred to PVDF membrane and probed with anti-Histone H3 (Unmodified Lys4) (0.1 μg/ml). Proteins were visualized using a goat-anti mouse secondary antibody conjugated to HRP (Cat. #AP124P) and a chemiluminescence detection system (Please see figures).
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Control:
Includes 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-Histone H3 (Unmodified Lys4) antibody and the Magna |