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    products/sample-preparation-kits/nuclear-extraction-kit-ab113474.pdf

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Nuclear Extraction Kit (ab113474)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (3)Q&A (30)References (150)

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Functional Studies - Nuclear Extraction Kit (ab113474)
  • Functional Studies - Nuclear Extraction Kit (ab113474)
  • Functional Studies - Nuclear Extraction Kit (ab113474)

Key features and details

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

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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.


    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.

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 100 tests
    1000X Protease Inhibitor Cocktail 1 x 110µl
    10X Pre-Extraction Buffer 1 x 10ml
    DTT Solution (1000X) 1 x 110µl
    ENE2 (Extraction Buffer) 1 x 10ml
  • Research areas

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

Associated products

  • Related Products

    • Bradford Reagent (ab119216)
    • ENE2 (Extraction Buffer) (ab203377)

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 two drugs refered to as Cilo (50 and 100 mg/kg; Cilo50and Cilo100), and Pio (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

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (150)

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

ab113474 has been referenced in 150 publications.

  • Zarei M  et al. Ginger and turmeric lipid-solubles attenuate heated oil-induced oxidative stress in the brain via the upregulation of NRF2 and improve cognitive function in rats. Metab Brain Dis 36:225-238 (2021). PubMed: 33170419
  • Aguiari P  et al. Persistent COUP-TFII expression underlies the myopathy and impaired muscle regeneration observed in resistance to thyroid hormone-alpha. Sci Rep 11:4601 (2021). PubMed: 33633251
  • Hu R  et al. Lactobacillus reuteri-derived extracellular vesicles maintain intestinal immune homeostasis against lipopolysaccharide-induced inflammatory responses in broilers. J Anim Sci Biotechnol 12:25 (2021). PubMed: 33593426
  • Kapetanou M  et al. FoxO1 Is a Novel Regulator of 20S Proteasome Subunits Expression and Activity. Front Cell Dev Biol 9:625715 (2021). PubMed: 33634126
  • Li L  et al. Sunitinib treatment promotes metastasis of drug-resistant renal cell carcinoma via TFE3 signaling pathway. Cell Death Dis 12:220 (2021). PubMed: 33637706
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 33 Abreviews or Q&A

Inhibition of skin carcinogenesis by suppression of NF-κB dependent ITGAV and TIMP-1 expression in IL-32γ overexpressed condition

Excellent Good 4/5 (Ease of Use)
Abreviews
Abreviews
Good nuclear extraction kit for tissues and cells
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 14 2019

Nuclear extraction from BV-2 cells

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
The protocol is easy to understand and nuclear extraction were performed as described by protocol. Loading control (Histone H1) were examined.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Ms. Ji Hye Han

Verified customer

Submitted Aug 06 2018

Nuclear Extraction from mouse hepatocytes treated with a DNA cross-linking drug

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
Nuclear fractions were performed as described by product datasheet. The target Rad17 and loading control were examined.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jun 28 2018

Question

can kit be used with frozen tissue samples

Read More

Abcam community

Verified customer

Asked on Feb 13 2013

Answer

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

Read More

Abcam Scientific Support

Answered on Feb 13 2013

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?

Read More

Abcam community

Verified customer

Asked on Jan 07 2016

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.

Read More

Sam Washer

Abcam Scientific Support

Answered on Jan 07 2016

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?

Read More

Abcam community

Verified customer

Asked on Jul 02 2014

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.

Read More

Jeremy Kasanov

Abcam Scientific Support

Answered on Jul 02 2014

Question

Hi, I recently purchased the Episeeker Nuclear Extract kit, and have a quick question about the procedure. I plan to use it with adipose tissue, and I'm wondering whether I could use a rotor-stator homogenizer, or if I need to use a dounce-style homogenizer. If I use a rotor-stator homogenizer on a slow speed, will the nuclear membranes remain intact?

Read More

Abcam community

Verified customer

Asked on Jun 14 2013

Answer

A rotor-stator homogenizer even at low speed may disrupt the nuclear membrane. We recommend using a Dounce homogenizer for isolating intact nuclei.

Read More

Tom Ruyle

Abcam Scientific Support

Answered on Jun 14 2013

Question

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


 

Read More

Abcam community

Verified customer

Asked on Jan 09 2017

Answer

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

Read More

Abcam Scientific Support

Answered on Jan 09 2017

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).

Read More

Abcam community

Verified customer

Asked on Oct 03 2016

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.

Read More

Abcam Scientific Support

Answered on Oct 03 2016

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!



------------------

Read More

Abcam community

Verified customer

Asked on Jun 29 2012

Answer

Thank you for your inquiry.

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

ab113474

https://www.abcam.com/Nuclear-Extraction-Kit-ab113474.html (or use the following: https://www.abcam.com/Nuclear-Extraction-Kit-ab113474.html).

ab113477

https://www.abcam.com/Nuclear-Nucleic-Acid-Free-Extraction-Kit-ab113477.html (or use the following: https://www.abcam.com/Nuclear-Nucleic-Acid-Free-Extraction-Kit-ab113477.html).

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/Chromatin-Extraction-Kit-ab117152.html (or use the following: https://www.abcam.com/Chromatin-Extraction-Kit-ab117152.html).

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.

Read More

Abcam Scientific Support

Answered on Jun 29 2012

1-10 of 33 Abreviews or Q&A

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