ABE435
Anti-trimethyl-Histone H3 (Lys36) Antibody
from rabbit, purified by affinity chromatography
Synonym(s):
Histone H3.1, Histone H3/a, Histone H3/b, Histone H3/c, Histone H3/d, Histone H3/f, Histone H3/h, Histone H3/I, Histone H3/j, Histone H3/k, Histone H3/l, H3K36me3
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All Photos(4)
About This Item
UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41
biological source
rabbit
Quality Level
antibody form
affinity isolated antibody
antibody product type
primary antibodies
clone
polyclonal
purified by
affinity chromatography
species reactivity
human, mouse
technique(s)
ChIP: suitable (ChIP-seq)
dot blot: suitable
immunocytochemistry: suitable
western blot: suitable
NCBI accession no.
UniProt accession no.
shipped in
wet ice
target post-translational modification
unmodified
Gene Information
human ... HIST1H3F(8968)
General description
Histones are nuclear proteins that form octameric structures which bind DNA to form units of chromatin called nucleosomes. The family of histones—H2A, H2B, H3, and H4—are key players in gene regulation. They undergo a number of post-translational modifications (PTM) in response to various stimuli including phosphorylation on serine and threonine residues and methylation on lysine residues. PTMs produce configural changes in histone proteins that may induce nucleosome remodeling and expose or hide DNA sequences from transcriptional complexes. Methylation occurs on histone H3 at lysine 4, 9, 27, 36, and 79 residues. Previous studies have reported that methylation of H3 lysine 9 provides a binding site for heterochromatin adaptor proteins (HP1) which are involved in the formation of heterochromatin and gene repression. Phosphorylation of histones may also produce similar repressive effects depending on the cellular context. Often, it is the pattern of different modifications on several histone residues that alter chromatin, rather than modification of single residues.
Specificity
Broad species reactivity expected due to similar sequence homology.
This antibody recognizes Histone H3 trimethylated at Lys36.
Immunogen
Epitope: Trimethylated Lys36
KLH-conjugated linear peptide corresponding to human Histone H3 trimethylated at Lys36.
Application
Anti-trimethyl-Histone H3 (Lys36) Antibody is a rabbit polyclonal antibody for detection of Histone H3 trimethylated at lysine 36. Also known as Anti-H3K36me3, this purified Ab is specificity verified by dot blot (DB) and has been validated in WB, ICC, DB, ChIP, ChIP-seq.
Dot Blot Specificity Analysis: Unmodified and various modified Histone peptides (see Table) were probed with Anti-trimethyl-Histone H3 (Lys36) (1:2,000 dilution).
Immunocytochemistry Analysis: A 1:500 dilution from a representative lot detected Histone H3 in A431 cells.
ChIP-Sequencing: Chromatin immunoprecipitation was performed using the Magna ChIP HiSens kit (cat#17-10460, 2 µg of anti-trimethyl-Histone H3 (Lys36) antibody(ABE435), 20 µL Protein A/G beads , and 1e6 crosslinked HeLa cell chromatin followed by DNA purification using magnetic beads. Libraries were prepared from Input and ChIP DNA samples using standard protocols with Illumina barcoded adapters, and analyzed on Illumina HiSeq instrument. An excess of eleven million reads from FastQ files were mapped using Bowtie (http://bowtie-bio.sourceforge.net/manual.shtml) following TagDust (http://genome.gsc.riken.jp/osc/english/dataresource/) tag removal. Peaks were identified using MACS (http://luelab.dfci.harvard.edu/MACS/), with peaks and reads visualized as a custom track in UCSC Genome Browser (http://genome.ucsc.edu) from BigWig and BED files.
Immunocytochemistry Analysis: A 1:500 dilution from a representative lot detected Histone H3 in A431 cells.
ChIP-Sequencing: Chromatin immunoprecipitation was performed using the Magna ChIP HiSens kit (cat#17-10460, 2 µg of anti-trimethyl-Histone H3 (Lys36) antibody(ABE435), 20 µL Protein A/G beads , and 1e6 crosslinked HeLa cell chromatin followed by DNA purification using magnetic beads. Libraries were prepared from Input and ChIP DNA samples using standard protocols with Illumina barcoded adapters, and analyzed on Illumina HiSeq instrument. An excess of eleven million reads from FastQ files were mapped using Bowtie (http://bowtie-bio.sourceforge.net/manual.shtml) following TagDust (http://genome.gsc.riken.jp/osc/english/dataresource/) tag removal. Peaks were identified using MACS (http://luelab.dfci.harvard.edu/MACS/), with peaks and reads visualized as a custom track in UCSC Genome Browser (http://genome.ucsc.edu) from BigWig and BED files.
Research Category
Epigenetics & Nuclear Function
Epigenetics & Nuclear Function
Research Sub Category
Histones
Histones
Quality
Evaluated by Western Blot in HeLa cell extract and recombinant Histone H3 protein.
Western Blot Analysis: A 1:500 dilution of this antibody detected Histone H3 on 10 µg of HeLa cell extract and recombinant Histone H3 protein.
Western Blot Analysis: A 1:500 dilution of this antibody detected Histone H3 on 10 µg of HeLa cell extract and recombinant Histone H3 protein.
Target description
~17 kDa observed. An uncharacterized band appears at ~52 kDa in some lysates.
Physical form
Affinity purified
Purified rabbit polyclonal in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Storage and Stability
Stable for 1 year at 2-8°C from date of receipt.
Analysis Note
Control
HeLa cell extract and recombinant Histone H3 protein
HeLa cell extract and recombinant Histone H3 protein
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
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Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 1
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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