Overview
50 preps, includes 50 ml cytoplasmic protein extraction buffer (CPEB), 50 ml protein solubilization buffer, 25 g protein solubilization buffer (PSB), 30 ml PSB diluent, 1 ReadyPrep 2-D Cleanup Kit (1632130), instruction manual; for 50 extractions of 50 mg of cells or tissue
Safety Data Sheet
Description
Description
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.
| ReadyPrep Kit for Fractionation by Cellular Location | Proteins Targeted | Catalog Number |
| ReadyPrep Protein Extraction Kit (Cytoplasmic/Nuclear) | Cytoplasmic and nuclear | 1632089 |
| ReadyPrep Protein Extraction Kit (Membrane I) | Membrane-anchored, with one or two transmembrane domains, and hydrophobic | 1632088 |
| ReadyPrep Protein Extraction Kit (Membrane II) | More than two transmembrane domains and/or proteins resistant to Membrane I Kit | 1632084 |
| ReadyPrep Protein Extraction Kit (Signal) | Associated with lipid rafts | 1632087 |
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