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Nuclear (Nucleic Acid-Free) Extraction Kit (ab113477)

Submit a review Q&A (7)References (3)

Key features and details

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

製品の概要

  • 製品名

    Nuclear (Nucleic Acid-Free) Extraction Kit

  • サンプルの種類

    Tissue, Adherent cells, Suspension cells

  • 全工程の試験時間

    1h 00m

  • 種交差性

    交差種: Mammals

  • 製品の概要

    Nuclear (Nucleic Acid-Free) Extraction Kit (ab113477) provides a simple and selective method for extracting nuclear proteins (nucleic-acid free) in just 1 hour for a variety of applications such as western blotting, protein-DNA binding assays, or any other procedures requiring optimized nucleic acid-free nuclear proteins.


    ab113477 can be used to extract nuclear proteins from mammalian cells and tissue samples free of nucleic acids attached.


    Not sure if this is the right product for you? Check out our Methods and tools to study histone modifications for help.

製品の特性

参考文献 (3)

ab113477 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab113477 は 3 報の論文で使用されています。

  • Gao A  et al. TAB182 aggravates progression of esophageal squamous cell carcinoma by enhancing β-catenin nuclear translocation through FHL2 dependent manner. Cell Death Dis 13:900 (2022). PubMed: 36289198
  • Dabrowska K  et al. The Role of Nrf2 Transcription Factor and Sp1-Nrf2 Protein Complex in Glutamine Transporter SN1 Regulation in Mouse Cortical Astrocytes Exposed to Ammonia. Int J Mol Sci 22:N/A (2021). PubMed: 34681893
  • Caropreso V  et al. Englerin A Inhibits EWS-FLI1 DNA Binding in Ewing Sarcoma Cells. J Biol Chem 291:10058-66 (2016). PubMed: 26961871

レビューと Q&A

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1-7 of 7 Abreviews or Q&A

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/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.

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Question

Hi,

I would like a kit which allows extract nuclear protein and more specially to extract chromatin binding protein.

Thanks you

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Answer

Thank you for your inquiry.

I am happy to confirm, that we do have several such kits.The classical kit for nuclear protein extraction is the ab113474. https://www.abcam.com/Nuclear-Extraction-Kit-ab113474.html (or use the following: https://www.abcam.com/Nuclear-Extraction-Kit-ab113474.html).I can however alsorecommend the kit ab113477 for example, which will allow extraction of all nuclear proteins in a nucleic acid free manner. Any enzymatic activity of a protein whould however been destroyed with this extraction kit. 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).The nuclear protein extraction kit will also extract DNA binding proteins, as illustrated withthe grpah where HDAC has been detected. For histone specific extraction however, we recommend the ab113476 kit. Indeed, Histones do bind even more strongly to the DNAhttps://www.abcam.com/Histone-Extraction-Kit-ab113476.html (or use the following: https://www.abcam.com/Histone-Extraction-Kit-ab113476.html).

I can stronglyrecommend you to look at our Guide, which will help you to choose the right kit for your application:
https://www.abcam.com/index.html?pageconfig=resource&rid=14321

I hope this information is helpful. Please do not hesitate to contact us again, should you have any further question.

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Question

I would like to isolate the nuclear protein with your Nuclear Extraction kit, ab113477, so my question is if I can get DNA-binding and nucleic acid-free extraction in a same time, I mean with the one kit . Thank you for you time.

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Answer

Thank you for contacting Abcam regarding ab113477.

This kit can be used to extract nucleic acid free nuclear proteins that can be used in protein-DNA binding assays.

I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions.

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Question

Would you be able to tell me if theAbcamEpiSeeker Nuclear (NA-free) Extraction Kit (cat #ab113477) can be used on frozen whole blood samples.

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Answer

Thank you for contacting us.

The lab sent the following advice:

Yes, it would be sufficient to simply spin these down and begin with the protocol for suspension cells. However, the nuclear extractionwill be in the red color, which may affect some of downstream application. The best way is to isolate the white blood cells for nuclear protein extraction. 1XNP1 buffer ishypotonic.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Question

Would you be able to tell me if theAbcamEpiSeeker Nuclear (NA-free) Extraction Kit (cat #ab113477) can be used on frozen whole blood samples.

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Answer

Thank you for contacting us.

The kit will be able to extract nuclear protein from intact nuclei of white blood cells but some percentage of the nuclei are likely to rupture when the samples are thawed. Have you tried thawing a sample to check if any cells are intact? You may find that nuclei are intact even if the outer plasma membrane has become damaged by the thaw. You will also want to separate the red blood cells from the white blood cells. This is usually accomplished with a Ficoll density gradient, but with fresh blood, not frozen/thawed blood. Have you tried the gradient separation, or any other method?

The protocol for suspension cells may be appropriate for your samples. The ENN1 buffer will lyse any remaining RBC as well as intact whole cells, without lysing the intact nuclei.

However, I have asked the laboratory that developed the kit to confirm that this is a valid protocol for your sample type, and I will send you their reply.

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Question

Many thanks for your mail and very useful information. I will be grateful if you can give me the discount code for ab193 antibody. We are working with brine shrimp (Artemia) and once I have the result I can send it to you. I am still looking for a good histone extraction method. I dont know which will suit the best for the antibody.I dont know if I can use any histone extraction kit. I will be grateful for your help. Thank you in advance Kind regards

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Answer

Thank you for your kind response. As I have explained in my previous e-mail any human lysate will probably have high levels of histone acetylation. However, it may be useful to treat the cells to maximize the level of acetylation. We have a very informative landing page for Epigenetics & Nuclear studies, please take a look at it: https://www.abcam.com/index.html?pageconfig=resource&rid=13907 Nuclear extraction kits: - ab113474 - ab113477 - ab117152 Histon extraction kit: - ab113476 DISCOUNT CODE: 100%ABR-XXXXXX Expiration date: XXXXXXX I am very pleased to hear you would like to accept our offer and test ab193 in brine shrimp (Artemia). This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for brine shrimp (Artemia) and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Question

Hi, I would like order the nuclear extraction kit. Here I have some questions. I would like the do nuclear extraction and I have done for several times without kit, just did it according a protocol. But there are some genomic DNA included in the nuclear extractor. And when I did the demethylation essay, there was no demethylation happened. I think it is the genomic DNA blocked the target methylated DNA demethylated but I am not sure. Now, I would like try it using the nuclear extraction, but I have several questions: 1) I found there are two nuclear extraction kits, one is regular and the other one is nucleic acid free, for my experiment, which one is better, or they are the same? I found the steps and regents of geting rid of DNA are added in the second kit, do they affect the following demethylation assay? 2) Generally, how much protein we can get from a plate of 2*106 cells when we do the nuclear extraction? Thank you. Best -- Yan Wang

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Answer

Thanks for your inquiry. 1. Regular nuclear extraction kit is better as nuclear proteins extracted with this kit can keep enzymes active compared to the kit for nucleic acid free extraction. We are not clear which demethylation assay is performed. If using our DNA demethylation kit , the genomic DNA in nuclear extracts should not affect the assay as the genomic DNA will not bind to the antibody or substrate coated on the plate.2. In general, 50-60 ug nuclear proteins can be yield from 2 million cells . I hope this was helpful. Please feel free to contact me if you require further assistance.

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