Thermo Scientific NE-PER Nuclear and Cytoplasmic Extraction Kit efficiently lyses cells and extracts individual cytoplasm in less than two hours and nuclear protein components.

NE-PER Nuclear and Cytoplasmic Extraction Kit Features:

•Fast — Nuclear and cytoplasmic fractions can be obtained in less than two hours
•Validated – NE-PER kits have been cited in over 950 reputable publications
•Universal – Nuclear proteins from cultured cells or tissues (only applicable for fresh samples)
•Expandable — Available in two kit formats for generating extracts from cells and tissues
•Convenient — Simple instructions , no gradient ultracentrifugation required
•Compatible — can be used for downstream assays including Western blotting, gel mobility assays, protein assays, reporter gene assays, and enzyme activity assays

The NE-PER kit is a nuclear protein extraction method that involves simple, stepwise lysis of cells and centrifugation to separate nuclear and cytoplasmic protein fractions. All you need is a benchtop microcentrifuge, test tubes, and pipette tools. NE-PER reagent efficiently solubilizes and separates various cytoplasmic and nuclear proteins into small aliquots with minimal cross-contamination or interference of genomic DNA and mRNA. After desalting or dilution, the isolated proteins can be used for immunoassays and protein interaction experiments such as mobility shift assays (EMSA), co-immunoprecipitation (Co-IP), and precipitation analyses.

A variety of methods are available for isolating cell nuclei and preparing nuclear protein extracts. Most methods for preparing nuclear extracts are tedious procedures requiring mechanical homogenization, freeze/thaw cycles, extensive centrifugation, or dialysis steps that may affect the integrity of nuclear proteins. The NE-PER Nuclear and Cytoplasmic Extraction Kit is a reagent-based protocol that progressively lyses cells, isolates cytoplasm from intact nuclei, and then extracts nuclear proteins from genomic DNA and mRNA. When working with cultured cells, this gentle process takes less than two hours and requires only a standard benchtop centrifuge. Additionally, active nuclear and cytoplasmic proteins can be recovered from the same cell culture or tissue sample.

This kit typically yields 200-500 µg of cytoplasmic protein and 100-200 µg of nuclear protein from 2 million cells (at a concentration of 1 mg/mL). Typical cross-contamination rates between the cytosolic and nuclear fractions are approximately 10%. This method primarily recovers soluble proteins from the nucleus rather than proteins bound in chromatin or integral membrane proteins of the nuclear envelope. The protein concentration of nuclear extracts can be easily controlled by using different volumes of nuclear extraction reagent (NER) in the extraction without any significant loss in extraction efficiency. The specific extraction of various nuclear proteins from cells is important for the success of many gene regulation studies. Although a variety of methods are available to isolate nuclei and prepare nuclear protein extracts, most of these methods are tedious and require mechanical homogenization, freeze/thaw cycles, and extensive centrifugation that may affect the integrity of many fragile nuclear proteins. or dialysis step. The NE-PER kit overcomes these issues and provides high-quality, concentrated nuclear protein extracts for EMSA and other analytical techniques.

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