Histone H3 (K36me3) polyclonal antibody
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Specification
Product Description
Rabbit polyclonal antibody raised against synthetic peptide of Histone H3 (K36me3).
Immunogen
A synthetic peptide (conjugated with KLH) corresponding to Histone H3, trimethylated at lysine 36.
Host
Rabbit
Reactivity
Human, Mouse
Form
Liquid
Recommend Usage
ELISA (1:100-500)
Western Blot (1:1000)
ChIP (5-10 ul/CHIP)
Dot Blot (1:100000)
The optimal working dilution should be determined by the end user.Storage Buffer
In whole antiserum (0.05% sodium azide).
Storage Instruction
Store at -20°C. For long term storage store at -80°C.
Aliquot to avoid repeated freezing and thawing.Note
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
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Applications
ChIP
ChIP assays were performed using human osteosarcoma (U2OS) cells. A titration of the antibody consisting of 2, 5, 10 and 15 ul per ChIP experiment was analysed. IgG (5 ug/IP) was used as a negative IP control. Quantitative PCR was performed using primer sets for the housekeeping gene GAPDH and for myogenic differentiation gene (MYOD). The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis.ChIP
ChIP was performed on sheared chromatin from 1 million HeLaS3 cells. IgG (2 ug/IP) was used as a negative IP control. The IP'd DNA was analysed by QPCR with optimized PCR primer pairs for the coding and promoter region of the active GAPDH gene, for the coding region of the inactive TSH2B gene and for the Sat2 satellite repeat.ChIP-Seq
The figure shows the results in 200 kb regions of chromosome 12 (including the GAPDH positive control), 6 and 7 and 14. These results clearly show an enrichment of the H3K36me3 at active genes.Western Blot (Cell lysate)
Western Blot (Cell lysate) analysis of 15 ug histone extracts of HeLa cells.Enzyme-linked Immunoabsorbent Assay
ELISA is a quantitative method used to determine the titer of the antibody using a serial dilution of antibody against Histone H3 (K36me3). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:12700.Dot Blot
Cross reactivity test using the Histone H3 (K36me3) antibody.
Dot Blot analysis was performed with peptides containing containing other modifications of histone H3.Other histone modifications include monomethylation and dimethylation of the same lysine and monomethylation, dimethylation and trimethylation of lysine 9, 27 and 79. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:100000. The figure shows a high specificity of the antibody for the modification of interest. -
Gene Info — HIST1H3A
Entrez GeneID
8350Protein Accession#
P68431Gene Name
HIST1H3A
Gene Alias
H3/A, H3FA
Gene Description
histone cluster 1, H3a
Omim ID
602810Gene Ontology
HyperlinkGene Summary
Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. This structure consists of approximately 146 bp of DNA wrapped around a nucleosome, an octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is found in the large histone gene cluster on chromosome 6p22-p21.3. [provided by RefSeq
Other Designations
H3 histone family, member A|histone 1, H3a
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Interactome
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Pathway
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Publication Reference
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The histone demethylase JMJD2A/KDM4A links ribosomal RNA transcription to nutrients and growth factors availability.
Salifou K, Ray S, Verrier L, Aguirrebengoa M, Trouche D, Panov KI, Vandromme M.
Nature Communications 2016 Jan; 7:10174.
Application:ChIP, Human, U-2 OS cells.
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The transcriptional and epigenomic foundations of ground state pluripotency.
Marks H, Kalkan T, Menafra R, Denissov S, Jones K, Hofemeister H, Nichols J, Kranz A, Stewart AF, Smith A, Stunnenberg HG.
Cell 2012 Apr; 149(3):590.
Application:ChIP-Seq, Mouse, Mouse embryonic stem cells.
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High-resolution analysis of epigenetic changes associated with X inactivation.
Marks H, Chow JC, Denissov S, Françoijs KJ, Brockdorff N, Heard E, Stunnenberg HG.
Genome Research 2009 Aug; 19(8):1361.
Application:ChIP-Seq, Mouse, Mouse embryonic stem cells.
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The histone demethylase JMJD2A/KDM4A links ribosomal RNA transcription to nutrients and growth factors availability.
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