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71183 NucBuster™ Protein Extraction Kit

71183
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71183-3
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      Glass bottle 100 rxns
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      Description
      OverviewThe NucBuster™ Protein Extraction Kit (Bruggink 2002) provides an alternative to the time intensive and cumbersome traditional methods for preparing nuclear extracts from mammalian cells. Time consuming traditional methods (up to 7 hours), based on a procedure originally described by Dignam et al. (1983), includes suspending cells in hypotonic solution, Dounce homogenization, centrifugation, and dialysis. The NucBuster Kit protocol is rapid and allows for the easy processing of multiple samples. The entire procedure from start to finish yields ready-to-use nuclear extract within 30 minutes. The composition of gentle detergents and salt in the final NucBuster extract is directly compatible with electrophoretic mobility shift assays (EMSA).

      The NucBuster protocol is based on two proprietary detergent-based solutions, NucBuster Extraction Reagent 1, optimized for cell lysis and removal of cytoplasmic components, and NucBusterExtraction Reagent 2, optimized for extraction of nuclear proteins. In addition, NucBuster extract is free of the 'stickiness' associated with release of genomic DNA, a problem associated with some traditional methods. No Dounce homogenization is required and there is no need for dialysis. The entire procedure is performed in a single microcentrifuge tube and requires only a vortex mixer and microcentrifuge. The kit provides enough reagents for 100 preparations of nuclear extract from 1 x 107 to 5 x 107 cells and the protocol is scalable.

      Catalogue Number71183
      Brand Family Novagen®
      Features and benefits
      • Nuclear extract preparation in 30 minutes
      • No homogenization or dialysis required
      • Procedure performed in a single microcentrifuge tube
      • Extracts suitable for activity assays, electrophoretic mobility shift assays (EMSA), and NoShift™ Transcription Factor Assays
      References
      ReferencesBruggink, F. and Hayes, S. 2002. inNovations 15, 9.
      Dignam, J.D., et al. 1983. Nucl. Acids Res. 11, 1475.
      Product Information
      2 × 7.5 mlNucBuster Extraction Reagent 1
      7.5 mlNucBuster Extraction Reagent 2
      100 µl100 mM DTT
      1 setProtease Inhibitor Cocktail Set I (lyophilized, makes 100 µl)
      Quality LevelMQ100
      Applications
      Biological Information
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Storage and Shipping Information
      Ship Code Shipped with Blue Ice or with Dry Ice
      Toxicity Multiple Toxicity Values, refer to MSDS
      Storage -20°C
      Do not freeze Ok to freeze
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalog Number GTIN
      71183-3 04055977254020

      Documentation

      NucBuster™ Protein Extraction Kit Certificates of Analysis

      TitleLot Number
      71183

      References

      Reference overview
      Bruggink, F. and Hayes, S. 2002. inNovations 15, 9.
      Dignam, J.D., et al. 1983. Nucl. Acids Res. 11, 1475.

      Citations

      Title
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    • Markus Bredel, et al. (2006) Tumor necrosis factor-α-induced protein 3 as a putative regulator of nuclear factor-κB-mediated resistance to O6-alkylating agents in human glioblastomas. Journal of Clinical Oncology 24, 274-287.
    • Ying Zhang, Mingjuan Liao and Maria L. Dufau. (2006) Phosphatidylinositol 3-kinase/protein kinase Cγ-induced phosphorylation of Sp1 and p107 repressor release have a critical role in histone deacetylase inhibitor-mediated depression of transcription of the luteinizing hormone receptor gene. Molecular and Cellular Biology 26, 6748-6761.
    • Susan M. Gordon, Noa Alon and Manuel Buchwald. (2005) FANCC, FANCE and FANCD2 form a ternary complex essential to the integrity of the Fanconi anemia DNA damage response pathway. Journal of Biological Chemistry 280, 36118-36125.
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    • Sridhar Mani, et al. (2005) Activation of the steroid and xenobiotic receptor (human pregnane X receptor) by nontaxane microtubule-stabilizing agents. Clinical Cancer Research 11, 6359-6369.
    • U. Panchapakesan, et al. (2005) PPARγ agonists exert antifibrotic effects in renal tubular cells exposed to high glucose. American Journal of Physiology: Renal 289, F1153-F1158.
    • Zachary T. Resch, et al. (2005) Stress-activated signaling pathways mediate the stimulation of pregnancy-associated plasma protein-A expression in cultured human fibroblasts. Endocrinology 147, 885-890.
    • Myriam Schaefer, et al. (2005) The transcription factors AP-1 and Ets are regulators of C3a receptor expression. Journal of Biological Chemistry 280, 42113-42123.
    • Ram N. Trivedi, et al. (2005) The role of base excision repair in the sensitivity and resistance to temozolomide-mediated cell death. Cancer Research 65, 6394-6400.
    • Qi Wu, et al. (2005) Mismatch repair participates in error-free processing of DNA interstrand crosslinks in human cells. European Molecular Biology Organization Reports 6, 551-557.
    • Jong In Yook, et al. (2005) Wnt-dependent regulation of the E-cadherin repressor snail. Journal of Biological Chemistry 280, 11740-11748.
    • Huie Jing, Jui-Hung Yen and Doina Ganea. (2004) A novel signaling pathway mediates the inhibition of CCL3/4 expression by prostaglandin E2. Journal of Biological Chemistry 279, 55176-55186.
    • Maoxiang Li, et al. (2003) The phosphatase MKP1 Is a transcriptional target of p53 involved in cell cycle regulation. Journal of Biological Chemistry 278, 41059-41068.
    • User Protocols

      Title
      TB338 NucBuster™ Extraction Reagent