Nuclear Extraction Kit

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Western Blot: Nuclear Extraction Kit [NBP2-29447] - 20 ug of HeLa cell proteins separated on 4-20% SDS-PAGE. Lane 1: Nuclear extract. Lane 2: Cytoplasmic extract. Lane 3: Total cell lysate.

Product Details

Summary
Kit Type
Extraction Kit

Order Details

Nuclear Extraction Kit Summary

Description
The Nuclear Extraction Kit provides a simple and convenient method for the isolation of nuclear and cytoplasmic extracts from mammalian cells and tissue samples. This procedure is relevant to the monitoring of translocation of cell signaling molecules from cytoplasm to the nucleus. Examples include translocation of NF-kB molecules to the nucleus in TNF-alpha treated cells, and translocation of mitogen-activated protein kinase to the nucleus in growth factor treated cells.

The Nuclear Extraction Kit can be used in the preparation of purified proteins for use in Western blotting, Eletrophoretic Mobility Assays (EMSA) and preparative purification of nuclear proteins.
Kit Type
Extraction Kit

Applications/Dilutions

Publications
Read Publications using
NBP2-29447 in the following applications:

Packaging, Storage & Formulations

Storage
Storage of components varies. See protocol for specific instructions.

Kit Components

Components
  1. 100X Protease Inhibitor Cocktail - PIC (100 uL)
  2. 10X PBS (2 x 50 mL)
  3. 10X Hypotonic Lysis Buffer (10 mL)
  4. 10% Detergent Solution (10 mL)
  5. 100 mM PMSF (10 mL)
  6. 1X Nuclear Extraction Buffer (10 mL)
  7. 1M DTT for nuclear extraction from tissue (100 uL)

Notes

Additional Equipment required, but not provided:
Teflon homogenizer (tissues)
Cell scraper (cells)
High-speed cold centrifuge and compatible centrifuge tubes
Microcentrifuge tubes
Deionized Water
Vortex

Alternate Names for Nuclear Extraction Kit

  • Nuclear Extraction Kit
  • Nuclear Extraction

Background

Nuclear Extraction

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Kits are guaranteed for 6 months from date of receipt.
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Publications for Nuclear Kit (NBP2-29447)(25)

We have publications tested in 1 confirmed species: Human.

We have publications tested in 2 applications: Cellular fractionation, WB.


Filter By Application
Cellular fractionation
(1)
WB
(4)
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Filter By Species
Human
(4)
All Species
Showing Publications 1 - 10 of 25. Show All 25 Publications.
Publications using NBP2-29447 Applications Species
Anti-inflammatory effects of astaxanthin in the human gingival keratinocyte line NDUSD-1 Miyachi M, Matsuno T, Asano K J Clin Biochem Nutr [PMID: 26060346]
ARAI M, TSUJI M, TSUCHIYA H. Protective Effects of Fucoidan Against Interleukin-1b-induced Inflammation in SW982 Human Synovial Cells. The Showa University Journal of Medical Sciences. 2014 Sep 27

Details:
Nuclear Extraction Kit used for isolation of nuclear fraction from SW982 cells (human synovial sarcoma cell line).
Awaji M, Saxena S, Wu L et al. CXCR2 signaling promotes secretory cancer-associated fibroblasts in pancreatic ductal adenocarcinoma FASEB J. May 26 2020 [PMID: 32453916]
Tsai TH, Yu CH, Chang YP et al. Protective Effect of Caffeic Acid Derivatives on tert-Butyl Hydroperoxide-Induced Oxidative Hepato-Toxicity and Mitochondrial Dysfunction in HepG2 Cells. Molecules Apr 28 2017 [PMID: 28452956] (Human) Human
Lee HK, Kim ID, Kim SW et al. Anti-inflammatory and anti-excitoxic effects of diethyl oxopropanamide, an ethyl pyruvate bioisoster, exert robust neuroprotective effects in the postischemic brain. Sci Rep. Feb 21 2017 [PMID: 28220827] (WB, Human) WB Human
Ramamoorthy H, Abraham P, Isaac B, Selvakumar D. Role for NF-kB inflammatory signalling pathway in tenofovir disoproxil fumarate (TDF) induced renal damage in rats Food Chem. Toxicol. Jan 1 2017 [PMID: 27899301] (WB) WB
Jiang H, He P, Xie J et al. Genetic deletion of TNFRII gene enhances the Alzheimer-like pathology in an APP transgenic mouse model via reduction of phosphorylated IkBalpha. Hum. Mol. Genet. 2014 May 13 [PMID: 24824215] (Cellular fractionation, WB, Human)

Details:
APP695 stably transfected 293 cells, Fig 7A. Nuclear and cytoplasmic fractions were probed with NF-kB, IkBa, or p-IkBa antibodies. PARP and beta actin antibodies were used as WB loading controls for the nuclear and cytoplasmic fractions, respectively.
Cellular fractionation, WB Human
Coleman SJ, Chioni AM, Ghallab M et al. Nuclear translocation of FGFR1 and FGF2 in pancreatic stellate cells facilitates pancreatic cancer cell invasion. EMBO Mol Med. 2014 Feb 6 [PMID: 24503018] (WB, Human)

Details:
WB & Cell Fractionation: pancreatic cancer cells cell lines, PS1 immortalized pancreatic cell, normal epithelial cells (Figs 2B, 2F, 2M, 3I, 4F). The fraction purity using the Nuclear Extraction kit confirmed with Lamin A/C and tubulin antibodies for the
WB Human
Kim SW, Kim HJ, Shin JH et al. Robust protective effects of a novel multimodal neuroprotectant oxopropanoyloxy benzoic acid (a salicylic acid/pyruvate ester) in the postischemic brain. Mol Pharmacol. 2011 Feb [PMID: 21036874]
Keiner B, Maenz B, Wagner R et al. Intracellular distribution of NS1 correlates with the infectivity and interferon antagonism of an avian influenza virus (H7N1). J Virol. 2010 Nov [PMID: 20844052]
Show All 25 Publications.

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Product General Protocols

View specific protocols for Nuclear Kit (NBP2-29447): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol

FAQs for Nuclear Kit (NBP2-29447). (Showing 1 - 1 of 1 FAQs).

  1. We are looking to isolate mtDNA that is free from nuclear DNA and nuclear DNA that is free from mtDNA. Do you know if these kits (NBP2-29447, NBP2-29448) will be able to do this? We are wanting the isolated DNA to use as a positive control for a copy number experiment.
    • You would need to use both kits in order to seperate mtDNA and nDNA. I was worried about the lysis buffer contained in both kits would force the DNA to precipitate, this can occur if the buffer is very basic or alcohol based. I also double checked to make sure all the buffers contained within the kit do not have any DNAses or RNAses present. Follow the protocol for both kits until you use the mitochondrial/nuclear lysis buffer and then proceed to collect the individual supernatants. I would proceed to use our Quik ChIP Kit. The way this kit works is by precipitating the DNA and then running it through a spin column included into the kit. Then the DNA is eluded of the column and the resulting product should be purified mDNA or nDNA depending on which supernatant you used. I reccomend on using a lot of starting material for your nuclear extract as the yield tends to be low, which is typical. I also reccomend to use 18 mega ohm DI water when starting your nuclear/mitochondrial extractions. The nuclear and mitochondrial extraction kits were designed for the purpose of extracting proteins and have not been tested for this particular application.

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