Overview

  • Product name
    Nuclear Extraction Kit
    See all Nuclear Extraction kits
  • Sample type
    Tissue, Adherent cells, Suspension cells
  • Assay time
    1h 00m
  • Product overview

    Nuclear Extraction Kit (ab113474) provides a simple and selective method along with all necessary reagents for nuclear protein extraction / nuclear protein fractionation in just 1 hour.


    The extracts can then be used in western blotting, protein-DNA binding assays, nuclear enzyme assays or any other procedures requiring optimized nuclear proteins. The protocol is fast and easy-to-use, and isolates very abundant yields of nuclear extract from mammalian cells or tissue samples.


    We recommend our alternative Soluble / Insoluble Nuclear Extraction kit to extract separate fractions of soluble and insoluble proteins.


    Not sure if this is the right product for you? Check out our EpiSeeker Sample Preparation Guide for help.


    Compared to other kits that use conventional nuclear extraction / nuclear fractionation methods, the buffers included in ab113474 contain much lower amounts of salts (80% less than conventional kits) and no SDS, which allows much better retention of enzyme activity in the nuclear extracts.

  • Notes

     

     

Properties

Images

  • Functional Studies - Nuclear Extraction Kit (ab113474)
    Functional Studies - Nuclear Extraction Kit (ab113474)
    Nuclear extracts were prepared from MCF-7 cells and the activity of HDACs were measured using different amounts of the extract. The result shown in the figure demonstrates the ab113474's high specificity.

Protocols

References

This product has been referenced in:
  • Parcon PA  et al. Apolipoprotein E4 inhibits autophagy gene products through direct, specific binding to CLEAR motifs. Alzheimers Dement 14:230-242 (2018). Read more (PubMed: 28945989) »
  • Celestini V  et al. Uncoupling FoxO3A mitochondrial and nuclear functions in cancer cells undergoing metabolic stress and chemotherapy. Cell Death Dis 9:231 (2018). Read more (PubMed: 29445193) »

See all 35 Publications for this product

Publishing research using ab113474? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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32 reviews or Q&A

10
Votes: Highest First

can kit be used with frozen tissue samples

Yes, this kit can be definitely used for frozen tissues. The lab confirmed it.

Dear professor!

I konw the Nuclear Extraction Kit which productde by Abcam.We wander obtain the cell nucleus,and then extrat the nuclear DNA.We want to know whether the kit can do it?

Thanks very much!



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Thank you for your inquiry.

We have two Nuclear Extraction Kits in our catalog at the moment:

ab113474

https://www.abcam.com/index.html?datasheet=113474 (or use the following: https://www.abcam.com/index.html?datasheet=113...

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Nuclear extraction from BV-2 cells

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
The protocol is easy to understand and nuclear extraction were performed as described by protocol. Loading control (Histone H1) were examined.
abreview image
Username

Ms. Ji Hye Han

Verified customer

Submitted Aug 06 2018

Nuclear Extraction from cisplatin treated mouse hepatocytes

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
Nuclear fractions were performed as described by product datasheet. The target Rad17 and loading control were examined.
abreview image
Username

Abcam user community

Verified customer

Submitted Jun 28 2018

Does the  cytosolic fraction include membrane protein as well. ( she will get the nuclear fraction againstthe rest of cell protein)?


Yes, the cytosolic fraction when using ab113474 Nuclear Extraction Kit includes membrane proteins.

Asking about the DTT concentration  ? ( someone told her the high DTT concentration may interrupt MS and co-IP)


Thank you for the follow up message.


I can confirm that the concentration of DTT included in the kit is 1M.


I hope this will be helpful. If you have any further questions, please do not hesitate to contact me.

I'm interested on the Nuclear Extraction Kit (ab113474). I would like to know if have you ever tried it with less amount of cells than what is written on the protocol booklet (around only half million cells).

Moreover, if after the extraction ...

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This kit will work better with 2 million cells however you can still use this with half a million cells, when using with 500,000 cells please keep the volume of buffers and reagents proportional to 1 million cells.

We recommend using Bradford ...

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ab113474
Tissue preparation section of protocol booklet states the following:
Tissue Samples
10.3.1 Weigh the tissue and cut it into small pieces. Place tissue
pieces in a clean homogenizer.
10.3.2 Add 5 mL of 1X Pre-Extraction Buff...

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I contacted the laboratories. They confirmed that PIC is not required for the tissue homogenization preparation protocol as the active proteases in tissue are much less than that in cultured cells.


Since we are concern about the nuclear integrity, we do not care too much about the cytosol. Therefore, DTT may can be excluded in Preextraction buffer too, because its main effect is to avoid the oxidation of the protein. Am I right?



I can confirm that if the cytosol is not required for much consideration, DTT can be excluded in pre-lysis buffer too.


I would like to use the HIF-1 alpha transcription factor assay kit (ab133104) but before I need to perform a nuclear extraction. I was looking in your website and found this product ab113474. Can I do a nuclear extraction using ab113474 and then ...

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Yes you can use ab113474, EpiSeeker Nuclear Extraction Kit, to purify soluble nuclear proteins from tissue culture cells or tissues and then detect HIF1 alpha in that extract using ab133104, HIF-1 alpha Transcription Factor Assay Kit.

1-10 of 32 Abreviews or Q&A

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