抗H3K27me3 (Ser28)抗体 | Anti-H3K27me3 (Ser28) 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

	ChIP assays were performed using HeLa cells treated with colcemid, the H3K27me3S28p antibody (bs-53053R) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 10,000 cells. A titration of the antibody consisting of 1, 5 and 10 μl per ChIP experiment was analysed. Additionally, ChIP was performed after incubation of the antibody with 5 nmol blocking peptide (cat. No. sp-091-050) for 1 hour at room temperature. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active genes c-fos and RPL30 and for the inactive 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).
	To determine the titer, an ELISA was performed using a serial dilution of the H3K27me3S28p antibody (bs-53053R). The antigen used was a peptide containing the histone modifications of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:8,300.
	A Dot Blot analysis was performed to test the cross reactivity of the H3K27me3S28p antibody (bs-53053R) with peptides containing other modifications of histone H3 and H4 and unmodified sequences from histone H3. One hundred to 0.2 pmol of the peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. The figure shows a high specificity of the antibody for the peptide containing the modifications of interest.
	Hela cells were stained with the H3K27me3S28p antibody (bs-53053R) and with DAPI. Cells were fixed with formaldehyde, permeabilized with sodium citrate and Triton X100 and blocked with PBS containing 2.5% BSA. Left side: cells were immunofluorescently labelled with the H3K27me3S28p antibody (diluted 1:200 and incubated for 1 hour at room temperature) followed by goat anti-rabbit antibody conjugated to DyLight 488. Right side: the nuclei were stained with DAPI, which specifically labels DNA. Phosphorylation of H3 on serine 28 is increased during late G2 phase and reaches a maximum in metaphase cells. This may explain the different staining intensities of different cells.
	HeLa cells were treated with colcemid to block the cell cycle in metaphase and were fixed with formaldehyde. Chromatin from 10,000 cells was sheared and used for immunoprecipitation (IP). IP was performed with 5 μl of the H3K27me3S28p antibody (bs-53053R). The immunoprecipitated proteins were analyzed by Western blot with the antibody diluted 1:500 in TBS-Tween containing 5% skimmed milk. Lane 1 shows the result of the IP; a negative IP control (no antibody added) and a positive control (sheared chromatin from 10,000 cells) are shown in lane 2 and 3, respectively.
	Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the H3K27me3S28p antibody (bs-53053R) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right. Lane 2 shows the result of the Western analysis with the antibody; lane 1 shows the same analysis after incubation of the antibody with 750 pmol blocking peptide for 1 hour at room temperature.

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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	H3K27me3 (Ser28) 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 histone H3 containing the trimethylated lysine 27 and the phosphorylated serine 28 (H3K27me3S28p), using a KLH-conjugated synthetic peptide
Purification	Whole antiserum from rabbit containing 0.05% azide

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