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ChIPAb+ Phospho-Histone H3 (Ser10) - ChIP Validated Antibody and Primer Set
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REFERENCES
Four enzymes cooperate to displace histone H1 during the first minute of hormonal gene activation.
Vicent, Guillermo Pablo, et al. (2011) Genes & development, (2011)
Species Reactivity Key Applications Host Format Antibody Type
Pl, Vrt WB, ChIP Mouse Protein G Purified Monoclonal Antibody
Description:
ChIPAb+ Phospho-Histone H3 (Ser10) - 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+ Phospho-Histone H3 (Ser10) set includes the Anti-phospho-Histone H3 (Ser10) antibody, a negative control antibody (purified Mouse IgG), and qPCR primers which amplify a 166 bp region within the promoter of the human GAPDH gene. The phospho-histone H3 (Ser10) and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of phospho-histone H3 (Ser10) associated chromatin.
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Specificity:
Recognizes histone H3, Mr 17 kDa, phosphorylated at Ser10.
Molecular Weight:
17 kDa
Epitope:
a.a. 1-19
Immunogen:
Immunogen was a synthetic peptide corresponding to amino acids 1-19 of histone H3, phosphorylated at Ser10.
Modifications:
Phosphorylation
Isotype:
IgG
Species Reactivity:
Green Plants
Vertebrates
Species Reactivity Note:
Human. The immunogen 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 untreated or colcemid-treated HeLa cells (2 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 2 μg of either a normal mouse IgG or Anti-phospho-Histone H3 (Ser10) antibody and the Magna ChIP G Kit (Cat. # 17-611). Successful immunoprecipitation of phospho-histone H3 (Ser10)-associated DNA fragments was verified by qPCR using Control Primers for untreated and treated chromatin samples (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. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
Western Blot Analysis:
Recombinant Histone H3 or acid extracts from colcemid-treated HeLa cells (1 μg) were resolved by electrophoresis, transferred to PVDF membrane and probed with 1 μg/mL anti-phospho Histone H3 (Ser10), clone CMA312. Proteins were visualized using goat anti-mouse secondary antibody conjugated to HRP 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 GAPDH promoter.
Quality Assurance:
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from colcemid-treated 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-phosph |