抗H3K4me3抗体 | Anti-H3K4me3 Antibody

• Bioss 社抗体製品の特長
• 価格表
• 使用例
• Background
• 製品情報

Bioss 社抗体製品の特長

• ウエスタンブロット、免疫組織化学（IHC-frozen、IHC-paraffin）、免疫蛍光（IF）、ELISAなどのアプリケーションに適した一次抗体がラインナップされています。
• 一次抗体はすべて、Protein AまたはProtein G精製されています。
• 幅広い研究分野に関連する抗体を取り揃えています。


※最新の適用、交差性情報の詳細は、下記製品情報項の「データシート」リンクをご参照ください。

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価格表

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使用例

	HeLa cells were stained with the Bioss antibody against H3K4me3 (Cat. No. bs-53103R) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K4me3 antibody (left) diluted 1:100 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
	Western blot was performed on whole cell (40 µg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells, and on 1 µg of recombinant histone H2A, H2B, H3.1, H3.3 and H4 (lane 3, 4, 5, 6 and 7, respectively) using the Bioss antibody against H3K4me3 (Cat. No. bs-53103R). The antibody was diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is shown on the right, the marker (in kDa) is shown on the left.
	A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K4me3 (Cat. No. bs-53103R) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:2,000. The figure shows a high specificity of the antibody for the modification of interest.
	ChIP was performed on sheared chromatin from 1 million HeLa cells using 1 µg of the Bioss antibody against H3K4me3 (Cat. No. bs-53103R) as described above. The IP'd DNA was subsequently analyzed on an Illumina Genome Analyzer. Library preparation, cluster generation, and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. The figure shows the peak distribution along the complete sequence and a 1.2 Mb region of the X-chromosome (figure A and B) and in two regions surrounding the GAPDH and EIF4A2 positive control genes, respectively (figure C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.
	ChIP assays were performed using human HeLa cells, the Bioss antibody against H3K4me3 (Cat. No. bs-53103R) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells. The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation (SNAP-ChIP K-MetStat Panel, Epicypher). A titration consisting of 1, 2, 5 and 10 µg of antibody per ChIP experiment was analyzed. IgG (2 µg/IP) was used as a negative IP control. Figure A. Quantitative PCR was performed with primers specific for the promoter of the active GAPDH and EIF4A2 genes, used as positive controls, and for the inactive MYOD1 gene, used as negative control. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes Figure B. Recovery of the nucleosomes carrying the H3K4me1, H3K4me2, H3K4me3 modifications and the unmodified H3K4 as determined by qPCR. The figure clearly shows the antibody is very specific in ChIP for the H3K4me3 modification. At higher concentrations, some H3K4me2 is also precipitated.

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Background

Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.

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製品情報

name	H3K4me3 Polyclonal Antibody
datasheet	最新のデータシートはこちらをご確認ください
Conjugation	Unconjugated
Gene ID	8350
Swiss Prot	P68431
Source	Polyclonal antibody raised in rabbit against the region of histone H3 containing the trimethylated lysine 4 (H3K4me3), using a KLH-conjugated synthetic peptide
Purification	Affinity purified polyclonal antibody in PBS containing 0.05% azide and 0.05% ProClin 300.

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