Histone Extraction Kit (ab113476)

Reviews (5)Q&A (25)References (30)

Overview

  • Product name
    Histone Extraction Kit
  • Sample type
    Tissue, Adherent cells, Suspension cells
  • Assay time
    1h 00m
  • Product overview

    Histone Extraction Kit (ab113476) is suitable for a quick isolation of total histone extracts from cells and tissue samples from mammals and other species.


    The amount of starting material can be as low as 105 cells or 1 mg of tissue. For best results, the cell number should be >106 cells or the tissue amount should be >10 mg.


    A total of 100 standard extractions (using 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.


     

  • Notes

    Histone purification is most often performed before the analysis of the global levels within a sample of post-translational histone modifications in western blot.

    Chromatin immunoprecipitation (ChIP) is the most common technique used for the analysis of histone modifications at specific sites within the genome. Use a chromatin extraction kit instead of a histone purification kit for the preparation of samples for ChIP.

Properties

Images

  • Functional Studies - Histone Extraction Kit (ab113476)
    Functional Studies - Histone Extraction Kit (ab113476)Nowacka-Woszuk, Joanna et al., PloS one?vol. 13,2 e0193464., Fig 4, doi:10.1371/journal.pone.0193464

    The differences in H3 acetylation level in fetal livers (female fetuses aged 19 dpc) from F1, F2, and F3 generations. measured with ab113476. Histones were prepared using ab115124
    Asterisk indicate a significant difference between the respective C and R groups: ∗∗ P < 0.01; NS not significant. Data presented as the means ± SE (n = 6 per group).

  • SDS-PAGE Analysis of samples prepared using Histone extraction Kit
    SDS-PAGE Analysis of samples prepared using Histone extraction Kit
  • Functional Studies - Histone Extraction Kit
    Functional Studies - Histone Extraction Kit

    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

  • Functional Studies - Histone Extraction Kit (ab113476)
    Functional Studies - Histone Extraction Kit (ab113476)
    Histone extracts were prepared from MCF-7 cells using the ab113476 and acetyl histone H3-K9 was quantified.

Protocols

References

This product has been referenced in:
  • Nowacka-Woszuk J  et al. Transgenerational effects of prenatal restricted diet on gene expression and histone modifications in the rat. PLoS One 13:e0193464 (2018). Read more (PubMed: 29474484) »
  • Wang J  et al. ATAC-Seq analysis reveals a widespread decrease of chromatin accessibility in age-related macular degeneration. Nat Commun 9:1364 (2018). Read more (PubMed: 29636475) »
See all 30 Publications for this product

Publishing research using ab113476? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 30 Abreviews or Q&A

Question

Please provide centrifugal force values for the RPM specifications in the the protocol. For example, 10k RPM = ? x g.
Does the purified product need to be de-salted before running on a gel?

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Answer

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.

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Histone Extraction Kit. Mosquito (Aedes spp).

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
This kit worked very well for extracting histones from the pharate larvae of Aedes mosquitoes, Since the pharate larvae are still inside the eggshell, this may have been a difficult sample.

To homogenize ~250 larvae (1 mg of eggs), I homogenized with a 1.5 ml plastic pestle in and then used a Biospec Bead Beater (Glen Mills) with Glass Beads (acid washed, 30-40 sieve) for 1.5 minutes. Proceeded with the rest of the protocol with no changes.

Marten Edwards

Verified customer

Submitted Nov 09 2018

Histone Extraction from Arabidopsis thaliana

 Good Good 4/5 (Ease of Use)
Abreviews
abreview image
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.

Jennifer Deke

Verified customer

Submitted 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.

Amy Cash

Verified customer

Submitted Aug 18 2017

Question

Dear technical support team: Why is so important to use this kit to isolation histone instead to use RIPA buffer? what is the difference?

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Answer

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.

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Histone Extraction from a non-model insect

 Good Good 4/5 (Ease of Use)
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.

Dr. Julie Reynolds

Verified customer

Submitted 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.

Abcam user community

Verified customer

Submitted Aug 20 2014

Question

Does this kit contain HDAC inhibitors?

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Answer

ab113476 does not contain any HDAC inhibitors.

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Question

I am interested in H1 histone extraction from human blood and skin and had problems with an in-house acid extraction protocol. I wonder how the EpiSeeker histone extraction kit performs on H1. Would it be able to extract H1 quantitatively from these tissues?

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Answer

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.

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Question

If I stored the cell pellets at -80oC, will the sample still be suitable for extracting nuclear protein and histone protein using the nuclear extraction kit (ab113474) and histone extraction kit (ab113476) respectively?

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Answer

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.

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

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