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    histone-extraction-kit-ab113476.pdf

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Histone Extraction Kit (ab113476)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (6)Q&A (25)References (52)

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Functional Studies - Histone Extraction Kit (ab113476)
  • SDS-PAGE Analysis of samples prepared using Histone extraction Kit
  • Functional Studies -  Histone Extraction Kit
  • Functional Studies - Histone Extraction Kit (ab113476)

Key features and details

  • Assay time: 1 hr
  • Sample type: Adherent cells, Suspension cells, Tissue

Overview

  • Product name

    Histone Extraction Kit
    See all Histone Extraction kits
  • 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

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 100 tests
    10X Pre-Lysis Buffer 1 x 10ml
    Balance Buffer 1 x 8ml
    DTT Solution 1 x 20µl
    Lysis Buffer 1 x 20ml
  • Research areas

    • Kits/ Lysates/ Other
    • Kits
    • Epigenetic kits
    • Sample Preparation
    • Epigenetics and Nuclear Signaling
    • Assays & Kits
    • Sample preparation

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
    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

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (52)

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

    ab113476 has been referenced in 52 publications.

    • Soofi A  et al. Regeneration after acute kidney injury requires PTIP-mediated epigenetic modifications. JCI Insight 5:N/A (2020). PubMed: 31917689
    • Li Q  et al. p53 Integrates Temporal WDR5 Inputs during Neuroectoderm and Mesoderm Differentiation of Mouse Embryonic Stem Cells. Cell Rep 30:465-480.e6 (2020). PubMed: 31940490
    • Sousounis K  et al. Eya2 promotes cell cycle progression by regulating DNA damage response during vertebrate limb regeneration. Elife 9:N/A (2020). PubMed: 32142407
    • Nam MH  et al. Aspirin inhibits TGFß2-induced epithelial to mesenchymal transition of lens epithelial cells: selective acetylation of K56 and K122 in histone H3. Biochem J 477:75-97 (2020). PubMed: 31815277
    • Tiwari SK  et al. Fatty acid ß-oxidation is required for the differentiation of larval hematopoietic progenitors in Drosophila. Elife 9:N/A (2020). PubMed: 32530419
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 31 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?

    Read More

    Abcam community

    Verified customer

    Asked on Sep 16 2013

    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.

    Read More

    Tom Ruyle

    Abcam Scientific Support

    Answered on Sep 16 2013

    Histone isolation from primary myoblasts

    Good Excellent 5/5 (Ease of Use)
    Abreviews
    Abreviews
    abreview image
    This is very good kit for isolating histones. Primary myoblasts isolated from the mouse were used for isolating histones. 0.25 million cells were used. Blots were developed using Histone H3 antibody (ab10799).

    Abcam user community

    Verified customer

    Submitted Jul 17 2019

    Histone Extraction Kit. Mosquito (Aedes spp).

    Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    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
    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
    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?

    Read More

    Abcam community

    Verified customer

    Asked on Mar 03 2015

    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.

    Read More

    Heather Allen

    Abcam Scientific Support

    Answered on Mar 03 2015

    Histone Extraction from a non-model insect

    Good Good 4/5 (Ease of Use)
    Abreviews
    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
    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?

    Read More

    Abcam community

    Verified customer

    Asked on Apr 21 2014

    Answer

    ab113476 does not contain any HDAC inhibitors.

    Read More

    Kevin Hanson

    Abcam Scientific Support

    Answered on Apr 21 2014

    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?

    Read More

    Abcam community

    Verified customer

    Asked on Jun 20 2013

    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.

    Read More

    Tanya Bagrij

    Abcam Scientific Support

    Answered on Jun 20 2013

    1-10 of 31 Abreviews or Q&A

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