CelLytic^™ NuCLEAR^™ Extraction Kit 

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A number of different procedures in the detailed technical bulletin enable the selection that best fits a particular application. For example, choose between detergent and non-detergent extraction of nuclear protein or between the standard hypotonic lysis buffer for most cell types and isotonic lysis buffer for fragile cells. In addition, the kit provides a procedure for salt reduction from the nuclear extract with dilution buffer. CelLytic-NuCLEAR offers the flexiblity you need for optimal protein extraction. Extracts can be prepared in less than 2 hours and are highly pure since there is little or no cross-contamination between nuclear and cytoplasmic extracts.

Within this kit is a complete system for preparing nuclear and cytoplasmic protein extracts from mammalian tissue or cultured cells. All reagents necessary for extraction are included.

CelLytic is a trademark of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co.

NuCLEAR is a trademark of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co.

1 kit sufficient for 10 extractions (1 ml packed cell volume)

1 kit sufficient for 100 extractions (100 μl packed cell volume)

dry ice

−20°C

Dyer, R.B. and Herzog, N.K., Isolation of intact nuclei for nuclear extract preparation from a fragile B-lymphocyte cell line. Biotechniques 19, 192-195, (1995) abstract

Lee, K.A., et al., A small-scale procedure for preparation of nuclear extracts that support efficient transcription and pre-mRNA splicing. Gene Anal. Tech. 5, 22-31, (1988) abstract

Dignam, J.D., et al., Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 11, 1475-1489, (1983) abstract

Guidez, F., et al., Differential utilization of Ras signaling pathways by macrophage colony-stimulating factor (CSF) and granulocyte-macrophage CSF receptors during macrophage differentiation. Mol. Cell. Biol. 18, 3851-3861, (1998) abstract

UN 3316 9/PG 3

10× Lysis Buffer, Hypotonic 7 mL

3× Dilution and Equilibration Buffer 90 mL

1 M DTT 0.4 mL

Extraction Buffer 10 mL

IGEPAL^® CA-630 10% Solution 4 mL

5× Lysis Buffer, Isotonic 14 mL

Protease Inhibitor Cocktail 1 mL P8340