NucleoSpin RNA/Protein—isolation and purification of RNA and protein from ONE lysate
NucleoSpin RNA/Protein is a complete mini kit with recombinant DNase and shredders for rapid, high-yield isolation and purification of total RNA and protein from cells and tissue. This kit allows simultaneous isolation of RNA and protein from a single lysate in one working procedure, from a wide variety of starting materials. The simple, fast, and environmentally friendly column-based procedure does not use phenol, chloroform, or acetone.
NucleoSpin RNA/Protein is a complete mini kit with recombinant DNase and shredders for rapid, high-yield isolation and purification of total RNA and protein from cells and tissue. This kit allows simultaneous isolation of RNA and protein from a single lysate in one working procedure, from a wide variety of starting materials. The simple, fast, and environmentally friendly column-based procedure does not use phenol, chloroform, or acetone.
This method is very useful for researchers interested in gene regulation mechanisms, such as knockdown of mRNA expression by siRNAs and its resulting influence on protein expression. Such studies are often complicated by small sample sizes and the need to use differing and often incompatible techniques to obtain RNA and protein. Dividing a sample into several parts for independent purification procedures is difficult, especially for small samples (e.g., biopsy material). Moreover, the isolation of RNA and protein from different sample aliquots usually necessitates proof of sample equality because the analysis might be mistrusted if the experimental samples used for nucleic acid and protein purification are not absolutely identical. NucleoSpin RNA/Protein resolves these issues by accommodating small sample sizes and providing compatible techniques for RNA and protein isolation.
Samples are lysed by incubation in a buffer containing large amounts of chaotropic salts. This not only destroys the cell structure of the sample, but simultaneously inactivates enzymes such as RNases, phosphatases, and proteases, and thus prevents RNAs and proteins from being degraded. The buffer dissolves even sparingly soluble proteins virtually quantitatively, and at the same time creates appropriate binding conditions for RNA adsorption to the silica membrane of RNA/Protein Columns. Thus, RNA and protein from the same sample can be separated by simply centrifuging the lysate through the NucleoSpin RNA/Protein spin column—the RNA is bound to the membrane while the protein is resolved in the column flowthrough.
Overview
- Complete mini kit with rDNase* and shredders
- High-quality RNA suitable for downstream applications
- High protein yield independent of protein size, localization, or modifications
- Simplifies transcriptional and translational gene expression profiling
- Easy protein quantification with the Protein Quantification Assay
* The amount of DNA contamination is significantly reduced during on-column rDNase digestion. However, in very sensitive applications it might be possible to detect traces of DNA. The NucleoSpin RNA/Protein system is checked by the following procedure: One million HeLa cells are subjected to RNA isolation according to the protocol. RNA eluate is used as template for PCR detection of a 1-kb fragment in a 30-cycle reaction. Generally, no PCR fragment is obtained if the rDNase is applied. However, a strong PCR fragment is obtained if rDNase is omitted. The eventuality of DNA detection with PCR increases with:
- The number of DNA copies per preparation:
Single copy target < plastidial/mitochondrial target < cells transfected with plasmid - Decreasing PCR amplicon size
More Information
| Technology | Silica membrane | |
| Format | Mini spin columns | |
| Starting material | <5 x 106 cultured cells <30 mg human/animal tissue 100 mg plant tissue |
|
| Total RNA | Total Protein | |
| Fragment size | 200 bp–20 kb | 15–200 kDa |
| Typical yield | <70 µg | <1,200 µg |
| A260/280 | 1.90–2.10 | Does not apply |
| Elution volume | 40–100 µl | 10–100 µl |
| Preparation time | 30 min/6 preps | 35 min/6 preps |
Applications
- Rapid isolation and purification of total RNA and protein from cultured cells and tissue
- Gene expression profiling, siRNA experiments, analysis of transgenic organisms, and drug screening
- Use for RT-PCR, 5' nuclease assays for probe-based qPCR, blotting, and microarrays, among others
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
NucleoSpin RNA/Protein application data
This product overview describes the use of the NucleoSpin RNA/Protein kit for the concurrent isolation of RNA and total cellular protein from undivided samples in one working procedure. Protein and RNA are obtained in separate fractions. This kit is especially valuable for unique, small, and precious samples. Isolated RNA is suitable for all common downstream applications and is of comparable high quality to RNA isolated with the proven NucleoSpin RNA kit. Isolated protein is immediately suitable for SDS-PAGE and Western blot analysis.
Learn more Other RNA purification kitsTakara Bio USA, Inc.
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