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ChIPAb+ Monomethyl-Histone H4 (Lys20)
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PRODUCT FAMILY INFORMATION
Histone H4 Antibodies and Proteins
Millipore’s Histone H4 antibodies demonstrate specificity against linker histone H4. See below for H4 Antibodies, ChipAb+ Primer sets, and peptides, based on the expertise of Upstate & Chemicon.
EMD Millipore’s Histone H4 antibody demonstrates specificity against linker histone H4. See below for data and references for EMD Millipore’s Histone H4 antibodies and proteins. All EMD Millipore antibodies and proteins are based on the expertise of Upstate & Chemicon.
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Species Reactivity Key Applications Host Format Antibody Type
H, Pl, Vrt WB, ChIP Rabbit Affinity Purified Polyclonal Antibody
Description:
ChIPAb+ Monomethyl-Histone H4 (Lys20) - 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+ Monomethyl-Histone H4 (Lys20) set includes the Anti-monomethyl-Histone H4 (Lys20) antibody, a negative control antibody (purified Rabbit IgG), and qPCR primers, which amplify a 212 base pair PCR product within the human GAPDH coding region. The monomethyl histone H4 (Lys20) and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of monomethyl-histone H4 (Lys20) associated chromatin.
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Specificity:
Recognizes histone H4, Mr ~11 kDa.
Molecular Weight:
~11 kDa
Epitope:
a.a. 15-24
Immunogen:
The immunogen was a synthetic peptide corresponding to amino acids 15-24 of histone H4, monomethylated on Lys20.
Modifications:
Methylation
Species Reactivity:
Human
Green Plants
Vertebrates
Species Reactivity Note:
Human. Not tested in other species. 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 HeLa cells (2 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 4 μg of either a negative control antibody or Anti-Monomethyl-Histone H4
(Lys20) antibody and the Magna ChIP A Kit (Cat. #17-610). Successful immunoprecipitation of monomethyl-histone H4 (Lys20)-associated DNA fragments was verified by qPCR using GAPDH coding region ChIP Primers versus Control Primers corresponding to the GAPDH promoter (Please see figures). Data is presented as percent input of each IP sample relative to input chromatin, with immunoprecipitated DNA from negative control antibody shown as (-) and monomethyl-histone H4 (Lys20) shown as (+).
Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
Western Blot Analysis:
0.1 μg/mL of proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures)
Peptide Blocking Assay:
40 μg of histone H4 peptide containing monomethyl Lys20 abolished detection of histone H4 by anti- |