Nuclear Extraction Kit

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.

• Kit Type: Extraction Kit

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

• Dilutions:
  • Western Blot

Packaging, Storage & Formulations

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

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.

Publications for Nuclear Kit (NBP2-29447)(28)

Publications using NBP2-29447

• Chen L, Dai M, Zuo W et al. NF-kB p65 and SETDB1 expedite lipopolysaccharide-induced intestinal inflammation in mice by inducing IRF7/NLR-dependent macrophage M1 polarization International immunopharmacology 2022-12-27 [PMID: 36580757]
• 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. 2014-09-27
  
  Details:
  Nuclear Extraction Kit used for isolation of nuclear fraction from SW982 cells (human synovial sarcoma cell line).
• Liang Y, Liu X, Zhou R et al. Chaetocin Promotes Osteogenic Differentiation via Modulating Wnt/Beta-Catenin Signaling in Mesenchymal Stem Cells Stem cells international 2021-02-06 [PMID: 33628276]
• Jong Ho Park, Amir H Ameri, Kaitlin E Dempsey, Danielle N Conrad, Marina Kem, Mari Mino-Kenudson, Shadmehr Demehri Nuclear IL-33/SMAD signaling axis promotes cancer development in chronic inflammation. The EMBO journal 2021-10-22 [PMID: 33616251]
• Awaji M, Saxena S, Wu L et al. CXCR2 signaling promotes secretory cancer-associated fibroblasts in pancreatic ductal adenocarcinoma FASEB J. 2020-05-26 [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 2017-04-28 [PMID: 28452956] (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. 2017-02-21 [PMID: 28220827] (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. 2017-01-01 [PMID: 27899301] (WB)
• 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]
• 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. 2014-05-13 [PMID: 24824215] (CellTox, 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.
• 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. 2014-02-06 [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
• 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. 2011-02-01 [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. 2010-11-01 [PMID: 20844052]
• Chaudhary P, Sharma R, Sharma A et al. Mechanisms of 4-hydroxy-2-nonenal induced pro- and anti-apoptotic signaling. Biochemistry. 2010-07-27 [PMID: 20565132]
• Li S, Banck M, Mujtaba S et al. p53-induced growth arrest is regulated by the mitochondrial SirT3 deacetylase. PLoS One. 2010-05-05 [PMID: 20463968]
• Parameswaran R, Muschen M, Kim YM et al. A functional receptor for B-cell-activating factor is expressed on human acute lymphoblastic leukemias. Cancer Res. 2010-06-01 [PMID: 20460528]
• Lau ST, Lin ZX, Leung PS. Role of reactive oxygen species in brucein D-mediated p38-mitogen-activated protein kinase and nuclear factor-kappaB signalling pathways in human pancreatic adenocarcinoma cells. Br J Cancer. 2010-02-02 [PMID: 20068565]
• Wang Z, Yao T, Song Z. Chronic alcohol consumption disrupted cholesterol homeostasis in rats: down-regulation of low-density lipoprotein receptor and enhancement of cholesterol biosynthesis pathway in the liver. Alcohol Clin Exp Res. 2010-03-01 [PMID: 20028367]
• Scharl M, Paul G, Weber A et al. Protection of epithelial barrier function by the Crohn's disease associated gene protein tyrosine phosphatase n2. Gastroenterology. 2009-12-01 [PMID: 19818778]
• Kim SW, Jeong JY, Kim HJ et al. Combination treatment with ethyl pyruvate and aspirin enhances neuroprotection in the postischemic brain. Neurotox Res. 2010-01-01 [PMID: 19636661]
• Nasir O, Wang K, Foller M et al. Relative resistance of SGK1 knockout mice against chemical carcinogenesis. IUBMB Life. 2009-07-01 [PMID: 19548318]
• Herve R, Schmitz T, Evain-Brion D et al. The PDE4 inhibitor rolipram prevents NF-kappaB binding activity and proinflammatory cytokine release in human chorionic cells. J Immunol. 2008-08-01 [PMID: 18641359]
• Sharma R, Sharma A, Dwivedi S et al. 4-Hydroxynonenal self-limits fas-mediated DISC-independent apoptosis by promoting export of Daxx from the nucleus to the cytosol and its binding to Fas. Biochemistry. 2008-01-08 [PMID: 18069800]
• Sampson VB, Rong NH, Han J et al. MicroRNA let-7a down-regulates MYC and reverts MYC-induced growth in Burkitt lymphoma cells. Cancer Res. 2007-10-15 [PMID: 17942906]
• Song Z, Deaciuc I, Zhou Z et al. Involvement of AMP-activated protein kinase in beneficial effects of betaine on high-sucrose diet-induced hepatic steatosis. Am J Physiol Gastrointest Liver Physiol. 2007-10-01 [PMID: 17702954]
• Vidaeff AC, Ramin SM, Gilstrap LC 3rd et al. Impact of progesterone on cytokine-stimulated nuclear factor-kappaB signaling in HeLa cells. J Matern Fetal Neonatal Med. 2007-01-01 [PMID: 17437195]
• He F, Li L, Kim D et al. Adenovirus-mediated expression of a dominant negative Ku70 fragment radiosensitizes human tumor cells under aerobic and hypoxic conditions. Cancer Res. 2007-01-15 [PMID: 17234773]
• Liu Z, Yang X, Tan F et al. Molecular cloning and characterization of human Castor, a novel human gene upregulated during cell differentiation. Biochem Biophys Res Commun. 2006-06-09 [PMID: 16631614]

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.