NE-PER™ Nuclear and Cytoplasmic Extraction Reagents
NE-PER™ Nuclear and Cytoplasmic Extraction Reagents
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Thermo Scientific™

NE-PER™ Nuclear and Cytoplasmic Extraction Reagents

The Thermo Scientific NE-PER Nuclear and Cytoplasmic Extraction Kit provides for efficient cell lysis and extraction of separate cytoplasmic andRead more
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Catalog NumberQuantity
7883575 mL
7883315 mL
Catalog number 78835
Price (USD)
778.65
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75 mL
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Price (USD)
778.65
Online Exclusive
866.00
Save 87.35 (10%)
Each
Add to cart
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The Thermo Scientific NE-PER Nuclear and Cytoplasmic Extraction Kit provides for efficient cell lysis and extraction of separate cytoplasmic and nuclear protein fractions in less than two hours.

Features of the NE-PER Nuclear and Cytoplasmic Extraction Kit:

Fast—obtain nuclear and cytoplasmic fractions of soluble proteins in less than two hours
Proven—the NE-PER Reagent Kit is referenced in more than 950 distinct publications
Versatile – nuclear protein from either cultured cells or tissues (intended for fresh samples only)
Scalable—two kit sizes for producing extracts from cells and tissues
Convenient—simple instructions do not require ultracentrifugation over gradients
Compatible—use for downstream assays, including Western blotting, gel-shift 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 centrifugal isolation of nuclear and cytoplasmic protein fractions. A benchtop microcentrifuge, tubes and pipettors are the only tools required. The NE-PER Reagents efficiently solubilize and separate cytoplasmic and nuclear proteins into fractions with minimal cross-contamination or interference from genomic DNA and mRNA. Once desalted or diluted, the isolated proteins can be used to perform immunoassays and protein interaction experiments, such as mobility shift assays (EMSA), co-immunoprecipitation (Co-IP) and pull-down assays.

There are a variety of methods available to isolate nuclei and prepare nuclear protein extracts. Most of the methods for preparing nuclear extracts are lengthy processes requiring mechanical homogenization, freeze/thaw cycles, extensive centrifugation or dialysis steps that may compromise the integrity of nuclear proteins. The NE-PER Nuclear and Cytoplasmic Extraction Reagent Kit is a reagent-based protocol that enables the stepwise lysis of cells, separation of the cytoplasm from the intact nuclei and then extraction of nuclear proteins away from genomic DNA and mRNA. This gentle process takes less than two hours and requires only a standard bench top centrifuge when using cultured cells. Furthermore, both active nuclear proteins and cytoplasmic proteins can be recovered from the same cell culture or tissue sample.

From two million cells, the kit typically yields 200 to 500 μg cytoplasmic protein and 100 to 200 μg nuclear protein (at a concentration of 1 mg/mL). Typical cross-contamination between cytosolic and nuclear fractions is about 10%. The method recovers primarily soluble proteins from the nucleus, not so much proteins bound in chromatin or integral membrane proteins of the nuclear envelope. The protein concentration of the nuclear extracts can be manipulated easily by varying the volume of nuclear extraction reagent (NER) used in the extraction without any significant loss in extraction efficiency. Specific extraction of nuclear proteins from cells is central to the success of many gene regulation studies. While a variety of methods exist to isolate nuclei and prepare nuclear protein extracts, most of these are lengthy processes requiring mechanical homogenization, freeze/thaw cycles, extensive centrifugation or dialysis steps that may compromise the integrity of many fragile nuclear proteins. The NE-PER Kit overcomes these problems, providing high-quality, concentrated nuclear protein extracts for use in EMSA and other analysis techniques.

Related Products
Subcellular Protein Fractionation Kit for Cultured Cells
Subcellular Protein Fractionation Kit for TissuesBenchtop Centrifuges
For Research Use Only. Not for use in diagnostic procedures.
Specifications
BufferLysis Buffer
Quantity75 mL
Reagent TypeCell Lysis Buffer, Detergent Solution, Subcellular Fractionation
Shipping ConditionAmbient
FormLiquid
Product LineNE-PER™
Product TypeNuclear and Cytoplasmic Extraction Reagent
Unit SizeEach
Contents & Storage
Upon receipt store kit components at 4°C. Product is shipped at ambient temperature.
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Frequently asked questions (FAQs)

How long does it take to complete the NE-PER Extraction reagents protocol?

Nuclear and cytoplasmic proteins can be isolated within 2 hours.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

After protein extraction using the NE-PER Extraction reagents, where is the genomic DNA located?

After extracting the nuclear proteins using the NER Reagent, genomic DNA will be located in the pellet.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Are there any considerations when adding protease inhibitors to the CER I and NER Reagents?

EDTA-free Halt Protease and/or Phosphatase Inhibitor Cocktails (e.g., Cat. No. 78425) can be added to CER I and NER reagents. Most commercially available protease inhibitor cocktails are also compatible; however, avoid protease inhibitors that contain alcohols. Because the concentration of the CER I and NER Reagents are critical, do not dilute these reagents by more than 5% when adding protease inhibitors.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Have the NE-PER Reagents been tested with plant tissue or yeast cells?

No. Also, nuclear protein was not successfully isolated from yeast cells that were pre-lysed with Y-PER Yeast Protein Extraction Reagent.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Is the final soluble nuclear fraction dialyzable when using the NE-PER Extraction reagents?

Yes. The nuclear fraction can be dialyzed in a Slide-A-Lyzer MINI Dialysis Unit (Cat. No. 69550) against a buffer, such as PBS, that is compatible with the specific downstream application. A buffer exchange can also be performed using the Thermo Scientific Protein Desalting Spin Columns (Cat. No. 89849) or Zeba Desalt Spin Columns (Cat. No. 89882).

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

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