製品の概要

  • 製品名
  • サンプルの種類
    Tissue, Adherent cells, Suspension cells
  • 全工程の試験時間
    1h 00m
  • 種交差性
    交差種: Mouse, Human, Drosophila melanogaster
    交差が予測される動物種: Mammal
  • 製品の概要

    Histone Extraction Kit (ab113476) is suitable for a quick preparation of total histone extracts from mammalian cells and tissue samples. The minimal amount of starting materials can be as low as 105 cells or 1 mg of tissue. For the best results, the cell number should be greater than 106 cells or the tissue amount should be greater than 10 mg. A total of 100 standard extractions (use 107 cells or 100 mg of tissue per extraction) can be performed with this kit. Yield of the total histone proteins can be up to 0.4 mg per 107 cells or per 100 mg of tissue. The yield may vary depending on the cell or tissue type.

  • 特記事項

    Shipped at +4°C. Store refrigerated at +4°C before preparation.

  • アプリケーション
    適用あり: Functional Studiesmore details

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab113476 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
Functional Studies Use at an assay dependent concentration. Total histone protein yield = 0.4mg/107 or 100mg tissue (yield may vary depending on cell or tissue type).

画像

  • Histones were purified from mouse liver cells using ab113476. Each lane was loaded with 15μg of purified histone.

    Image courtesy of Kevin Dalgaard, MPI for Immunobiology & Epigenetics, Freiburg, DE

  • Histone extracts were prepared from MCF-7 cells using the ab113476 and acetyl histone H3-K9 was quantified.

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参考文献

This product has been referenced in:
  • Alzrigat M  et al. EZH2 inhibition in multiple myeloma downregulates myeloma associated oncogenes and upregulates microRNAs with potential tumor suppressor functions. Oncotarget 8:10213-10224 (2017). WB ; Human . Read more (PubMed: 28052011) »
  • Gursoy-Yuzugullu O  et al. Epigenetic therapy with inhibitors of histone methylation suppresses DNA damage signaling and increases glioma cell radiosensitivity. Oncotarget 8:24518-24532 (2017). Read more (PubMed: 28445939) »

See all 16 Publications for this product

レビューと Q&A

The equivalent G Force values are:

1000 RPM = 110G
3000 RPM = 1000G
10,000 RPM = 8000G
12,000 RPM = 11,000G

No special preparation such as desalting needs to be done before loading the sample into the gel.

Abreviews
The Histone Extraction Kit (ab113476) was used to extract histones from adult A. thaliana leaves.

2.5 g leaf material was frozen and ground in liquid N2 and used for the extraction with 8 ml 1x Pre-Lysis Buffer. Two filtration steps through 100 μm and 30 μm nylon meshes and additional Pre-Lysis Buffer wash steps were performed in order to remove residual plant material.
The histone extract was precipitated with 4% perchloric acid o/n.

The Histone Extraction Kit is easy to use and generally works with A. thaliana leaves. However, the resulting sample contained some unexpected proteins, e.g. chloroplastic proteins, in addition to histones.
Username

Jennifer Deke

Verified customer

投稿 Sep 28 2017

Histone Extraction from a Honey Bee Brain

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Our lab used the Histone Extraction Kit (ab113476) to extract histones from brains of the Western honeybee, Apis mellifera.

A single brain (approximately 10 mg) was used for each sample. We quantified the histone extracts using absorbance at 280 nm on a NanoDrop 1000 spectrophotometer. Each extraction yielded 20-40 ug of total histone protein. We then used the extracted histones in the Histone H3 Total Acetylation Detection Fast Kit (Fluorometric) (ab131561).

The kit is very easy to use and we will use it again in the future.
Username

Amy Cash

Verified customer

投稿 Aug 18 2017

RIPA buffer can be used for extracting nuclear proteins but ab113476 is specifically for histone proteins. In general, the isolated histone protein concentration is much higher with use of ab113476than with RIPA buffer.

Abreviews
The EpiSeeker Histone Extraction kit was used to isolate histone proteins for use with the EpiSeeker Histone H3 Total Acetylation Detection Fast Kit. Histones were extracted pupae of the flesh fly, Sarcophaga bullata, which is a non-model insect.

The kit was easy and fast to use. It was easy to scale the reaction size to the amount of tissue.
Username

Dr. Julie Reynolds

Verified customer

投稿 Aug 21 2014

Histone Extraction from a non-model insect

Good Excellent 5/5 (Ease of Use)
Abreviews
This kit was used to isolate histones from pupae of the fleshy fly, Sarcophaga bullata, a non-model insect. A single pupa, weighing approximately 100 mg, was used in each sample. Each extraction yielded 14-19 ug/ul of histone protein.
The kit was easy to use and simple to scale to the size of my insects. The extracted histones worked well with the EipSeeker Histone H3 Total Acetylation Detection Fast Kit.
Username

Abcam user community

Verified customer

投稿 Aug 20 2014

ab113476 does not contain any HDAC inhibitors.

I can confirm that ab113476: EpiSeeker Histone Extraction Kit would be suitable to use for human blood and skin and would extract total histone proteins, including H1, that would be quantifiable.

Nuclear Protein extraction and Histone protein extraction from cell samples frozen at -80oC could still work using ab113474 and ab113476. However, the enzyme activity of nuclear extracts from frozen tissues may be much lower than that from fresh tissues.

Ab113476 can be used to extract histones from a protozoan parasite if you use the protocol for histone extraction from tissues.

1-10 of 29 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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