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ChIPAb+ Histone H2B - ChIP Validated Antibody and Primer Set
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PRODUCT FAMILY INFORMATION
Histone H2B Antibodies & Proteins
Millipore’s Histone H2B antibodies demonstrate specificity against histone H2B. See below for acetyl-, methyl-, phospho- H2B Antibodies and Proteins, based on the expertise of Upstate & Chemicon.
EMD Millipore’s Histone H2B antibodies and proteins are based on the expertise of Upstate & Chemicon. All have been well validated and published. See the ordering tab for a comprehensive list of our Histone H2B products and direct links to their technical pages for data, references and related products.
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Description:
ChIPAb+ Histone H2B - ChIP Validated Antibody and Primer Set
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 H2B set includes the Histone H2B antibody, a negative control rabbit IgG, and qPCR primers which amplify a 110 bp region of human β-globin promoter. The Histone H2B and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Histone H2B-associated chromatin.
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Specificity:
Recognizes and is specific for Histone H2B, MW ~15 kDa. An additional band at ~23 kDa is detected in some samples, which likely corresponds to ubiquityl-Histone H2B.
Molecular Weight:
~15 kDa
Immunogen:
KLH-conjugated, synthetic peptide corresponding to amino acids 118-126 (CG-AVTKYTSSK) of human Histone H2B, with an N-terminal CG added for conjugation purposes.
Background Information:
Histone H2B is one of the 5 main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N terminal tail H2A is involved with the structure of the nucleosomes of the 'beads on a string' structure.
Species Reactivity:
Chicken
Xenopus
Human
Yeast (S. cerevisiae)
Species Reactivity Note:
Human, chicken, yeast and Xenopus. Others not tested. Broad species cross-reactivity expected based on 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 either 1 µL of Normal Rabbit IgG or 1 µL of Anti-Histone H2B and the Magna ChIP™ A Kit (Cat. # 17-610).
Successful immunoprecipitation of Histone H2B associated DNA fragments was verified by qPCR using ChIP Primers, human β-globin for a positive, and human GAPDH primers as a negative assay (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.
Recombinant Xenopus Histone H2B, chicken core histones and acid extract from either sodium butyrate or colcemid treated HeLa cells were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Histone H2B (1:5,000 dilution).
Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and |