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ChIPAb+ Histone H3 (CT) - ChIP Validated Antibody and Primer Set, rabbit monoclonal
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Species Reactivity Key Applications Host Format Antibody Type
H, M, R, Yeast (S. cerevisiae), Ch WB, IP, DB, ChIP Rabbit Culture Supernatant Monoclonal Antibody
Description:
ChIPAb+ Histone H3 (CT) - ChIP Validated Antibody and Primer Set, rabbit monoclonal
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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 (C-term) set includes the Histone H3 (C-term) antibody, a negative control supernatant (rabbit), and qPCR primers which amplify a 110 bp region of human β-Globin. The Histone H3 (C-term) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Histone H3-associated chromatin.
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Specificity:
Recognizes Histone H3 near the C-terminus.
Molecular Weight:
~17 kda
Epitope:
C-Terminus
Immunogen:
KLH-conjugated, synthetic peptide corres-ponding to the C-terminus of human Histone H3.
Isotype:
IgG
Background Information:
Histone H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3 is involved with the structure of the nucleosomes of the 'beads on a string' structure. Histone proteins are highly post-translationally modified, and Histone H3 is the most extensively modified of the five histones. The term "Histone H3" alone is purposely ambiguous in that it does not distinguish between sequence variants or modification state. Histone H3 is an important protein in the emerging field of epigenetics, where its sequence variants and variable modification states are thought to play a role in the dynamic and long term regulation of genes.
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Species Reactivity:
Human
Mouse
Rat
Yeast (S. cerevisiae)
Chicken
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Species Reactivity Note:
Human, mouse, rat, yeast and chicken. Broad species cross-reactivity expected due to sequence homology.
Application Notes:
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 4 µL of a negative control supernatant, or 4 µL of Anti-Histone H3 (C-term) and the Magna ChIP™ A Kit (Cat. # 17-610).
Successful immunoprecipitation of Histone H3 associated DNA fragments was verified by qPCR using ChIP Primers, β-Globin as well as GAPDH promoter 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 ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
Western Blot Analysis:
Representative Lot Data.
Acid extracts from colcemid treated HeLa cells were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Histone H3 (C-term) (1:10 |