Nuclear Extraction Kit (ab113474)

Reviews (2)Q&A (30)References (36)

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)Ragab, Diaa et al., PloS one?vol. 9,5 e95313., Fig 6, doi:10.1371/journal.pone.0095313

    Effect of cilostazol (50 and 100 mg/kg; Cilo50and Cilo100), pioglitazone (3 and 10 mg/kg; Pio3 and Pio10), and their combination (Cilo50and Pio3) on the PPAR-γ transcription activity in rats subjected to ischemia (45 min)/reperfusion (24 hrs).
    Drugs were administered orally for 14 days then subjected to ischemia/reperfusion. Values are expressed as mean ± S.E.M (n = 6). Data are compared with sham operated control (#), I/R control (∗), Cilo50 (), Pio3 (‡), and combination (§) pretreated groups (one-way ANOVA followed by Tukey Multiple Comparison Test) at P<0.05.

  • Functional Studies - Nuclear Extraction Kit (ab113474)
    Functional Studies - Nuclear Extraction Kit (ab113474)Park, Jisu et al., PloS one?vol. 10,11 e0141988., Fig 4, doi:10.1371/journal.pone.0141988
    B16F10 cells were treated with 30 µM of DMPB for the indicated time periods. Cytoplasmic and nuclear fractions were isolated and analyzed by Western blotting.
  • 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 36 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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1-10 of 32 Abreviews or Q&A

Question

can kit be used with frozen tissue samples

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Answer

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

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Question

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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Answer

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=113474).

ab113477

https://www.abcam.com/index.html?datasheet=113477 (or use the following: https://www.abcam.com/index.html?datasheet=113477).

Both kits are intended for the extraction of protein from the nucleus.

ab113477 is designed to extract Proteins nucleic acid free.

We also have ab117152 (EpiSeeker Chromatin Extraction Kit) that will extract DNA and theDNA bound chromatin.

https://www.abcam.com/index.html?datasheet=117152 (or use the following: https://www.abcam.com/index.html?datasheet=117152).

I am sorry to confirm that we do not have kits specifically for protein free DNAextraction.

I would also like to make you aware that we do have scientific support in Chinese. If you prefer to speak to us in Chinese please write to mailto:hk.technical@abcam.com.

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Nuclear extraction from BV-2 cells

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

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
abreview image
Nuclear fractions were performed as described by product datasheet. The target Rad17 and loading control were examined.

Abcam user community

Verified customer

Submitted Jun 28 2018

Question

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


 

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Answer

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

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Question

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


 

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Answer

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.

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Question

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 it is possible to quantify the protein amount with the Pierce BCA assay (if any component of the pre-extraction buffer or of the extraction buffer can interfere).

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Answer

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 for quantification, but the BCA Pierce assay will also be suitable.

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Question

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 Buffer containing 5 μL of
DTT Solution per gram of tissue, and homogenize tissue
pieces (50-60 strokes).
10.3.3 Incubate on ice for 15 minutes and centrifuge for
10 minutes at 12,000 rpm at 4°C.
10.3.4 Remove the supernatant.

This does not mention PIC. Do we need to add this? Do they use stock solution of the pre extractio buffer, or do they use pre extraction buffer as prepared earlier in the protocol with DTT and PIC already added?

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Answer



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.

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Question


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?

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Answer



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

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Question


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 do my experiment with ab133104?

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Answer



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.

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1-10 of 32 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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