Description
Application
CHEMICON′s Nuclear Extraction Kit (Catalog No. 2900) provides a simple and convenient method for the isolation of cytoplasmic and nuclear samples from mammalian cell culture or tissue samples. The Nuclear Extraction Kit can be used in the preparation of purified proteins for use in Western blotting, Electrophoretic Mobility Shift Assays (EMSA), and in CHEMICON′s Transcription Factor Assay product line.
For Research Use Only; Not for use in diagnostic procedures.
Research Category
All
Preparation Note
The Nuclear Extraction Kit is shipped and stored at -20ºC.
·Cytoplasmic Lysis Buffer (10x), Nuclear Extraction Buffer (1x), 10% Detergent, and the PBS Packets can be stored at 2-8ºC.
·DTT, 1M and Protease Inhibitor Cocktail must be stored at -20ºC. Avoid repeated freeze-thaw cycles.
Other Notes
Cytoplasmic Lysis Buffer, 10x: - (Part No. 90497) One vial containing 10mL of a concentrated lysis buffer. Dilute to a 1x in deionized water.
Nuclear Extraction Buffer: - (Part No. 90498) One vial containing 50mL of a 1x nuclear extraction buffer.
PBS Packets: - (Part No. 60093) Two pouches containing enough dry reagent to prepare 1 liter of a 1x PBS solution per pouch. Dilute packet in deionized water.
DTT, 1M: - (Part No. 90499) One vial containing 100 μL of 1M Dithiothreitol. Prior to use, dilute to a final concentration of 0.5mM (1:2000) in 1x Cytoplasmic Lysis Buffer and 1x Nuclear Extraction Buffer.
Protease Inhibitor Cocktail: - (Part No. 90492) One vial containing 100μL of protease inhibitors in DMSO for use with mammalian cell and tissue extract buffers. A mixture of protease inhibitors with broad specificity for the inhibition of serine, cysteine and aspartic acid proteases and aminopeptidases. Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF), pepstatin A, E-64, bestatin, leupeptin, and aprotinin. Contains no metal chelators (e.g. EDTA, EGTA). Prior to use, dilute 1/1000 in 1x Cytoplasmic Lysis Buffer and 1x Nuclear Extraction Buffer.
Detergent, 10%: - (Part No. 90500) One vial containing 3mL of 10% IGEPAL CA-630 (detergent). Cytoplasmic Lysis Buffer and Nuclear Extraction Buffer already contain detergent, however, under certain conditions, more detergent may be required - refer to Extraction Procedure.
Legal Information
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
