Use the ReadyPrep Protein Extraction Kit (Cytoplasmic/Nuclear) to enrich for both cytoplasmic and nuclear proteins. The kit separates cytoplasmic proteins from intact nuclei and then solubilizes nuclei.

• Employs a specially formulated buffer and differential centrifugation to isolate intact nuclei
• Contains a strongly chaotropic extraction buffer to efficiently solubilize nuclear proteins
• Includes the components of the ReadyPrep 2-D Cleanup Kit to remove extraction buffer that may interfere with IEF
• Extracted proteins are suitable for IEF, SDS-PAGE, and western blotting

The ReadyPrep Protein Extraction Kit (Cytoplasmic/Nuclear) uses a buffer/differential centrifugation protocol to generate enriched cytoplasmic fractions and intact nuclei (Dignam et al. 1983, Zerivitz and Akusjarvi 1989). The nuclei are treated with a strongly chaotropic extraction buffer to efficiently solubilize the nuclear proteins. The cytoplasmic fraction is processed with the included ReadyPrep 2-D Cleanup Kit to remove IEF-inhibiting compounds.  Although this kit is directed towards the preparation of nuclear and cytoplasmic protein samples for 2-D gel electrophoresis, the fractions are suitable for SDS-PAGE, western blotting, and other applications.

Kit Components

For 50 extractions of 50 mg of cells or tissue:

• Cytoplasmic protein extraction buffer (CPEB), 50 ml
• Protein solubilization buffer (PSB), 25 g
• 3PSB diluent, 30 ml
• ReadyPrep 2-D Cleanup Kit (1632130)
• Instruction manual

Bio-Rad offers a range of ReadyPrep Kits for Fractionation by Cellular Location. These kits reduce sample complexity, increase detection and identification of low-abundance proteins, and simplify proteomic studies.

Related Products

• ReadyPrep Kits for Fractionation by Solubility
• Aurum™ Ion Exchange Mini Kits for Fractionation by Charge
• Rotofor^® Liquid-Phase IEF Cells for Fractionation by pI
• Model 491 Prep Cell and Mini Prep Cell for Fractionation by Solubility
• ReadyPrep™ 2-D Starter Kit
• PROTEAN^® i12™ IEF System
• ReadyStrip™ IPG Strips
• ReadyPrep Reduction-Alkylation Kit (1632090)
• PowerPac™ HV High-Voltage Power Supply

Related Categories

• Protein Gels
• Electrophoresis Chambers
• 2-D Electrophoresis Buffers and Reagents
• Ampholytes
• Tube Gel IEF Systems
• Gel Imaging Systems
• 2-D Electrophoresis Workflow

Supporting Documentation

• ReadyPrep Protein Extraction Kit (Cytoplasmic Nuclear) Instruction Manual
• Sample Preparation: Tools for Protein Sample Extraction, Cleanup, Fractionation, and Depletion Brochure
• ReadyStrip IPG Strip Instruction Manual
• PROTEAN i12 IEF System Brochure
• Host Cell Protein (HCP) Workflow Quick Guide

References

Dignam JD et al. (1983). Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acid Res 11, 1475–1489. PMID: 6828386
Zerivitz K and Akusjarvi G (1989). An improved nuclear extract preparation method. Gene Anal Tech 6, 101–109. PMID: 2555292