概述

  • 产品名称
    核Extraction试剂盒
    参阅全部 Nuclear Extraction 试剂盒
  • 样品类型
    Tissue, Adherent cells, Suspension cells
  • 检测时间
    1h 00m
  • 种属反应性
    与反应: Mouse, Human
    预测可用于: Mammal
  • 产品概述

    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 for a variety of applications such as 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.

  • 说明

     

     

  • 经测试应用
    适用于: Functional Studiesmore details

性能

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应用

Our Abpromise guarantee covers the use of ab113474 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
Functional Studies Use at an assay dependent concentration.

图片

  • 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.

实验方案

文献

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 33 Publications for this product

发表研究结果有使用 ab113474？请让我们知道，以便我们可以引用本数据表中的参考文章。

客户评价及客户问答

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30 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

http://www.abcam.com/index.html?datasheet=113474 (or use the following: http://www.abcam.com/index.html?datasheet=11347...

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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.

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 homoge...

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

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?

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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