| NucleoSpin® RNA/Protein |
Total RNA and protein isolation
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| Features |
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| Parallel isolation of RNA and protein from unsplit samples |
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 • Complete mini kit with recombinant DNase and shredders for DNA-free* RNA
of high quality
• Reliable interpretation of RNA and protein levels
• High RNA yield and purity
• High protein yield independent of protein size, localization, modification, etc.
• Protein quantification possible by using the Protein Quantification Assay
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| Technology |
Silica-membrane technology
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| Format |
Mini spin columns
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| Sample material |
< 5 x 106 cultured cells
< 30 mg human/animal tissue
< 100 mg plant tissue
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Total RNA
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Total protein
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| Fragment size |
200 b - 20 kb
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15 - 300 kDa
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| Typical yield |
< 70 µg
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< 1200 µg
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| A260/280 |
1.9-2.1
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Does not apply
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Elution volume (RNA)
Resolubilization volume (protein) |
40 - 100 µl
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10 - 100 µl
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| Preparation time |
30 min/6 preps
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35 min/6 preps
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| Binding capacity |
200 µg
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Does not apply
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Applications**
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• Rapid purification of total RNA and protein from cultured cells and tissue
• Gene expression profiling, siRNA experiments, analysis of transgenic organisms, drug screening
• Broad spectrum of starting material tested and proteins already detected |
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| Starting materials successfully used |
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Proteins successfully analyzed |
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In general: cultured cells / fresh tissue /frozen tissue / tissue in RNAlater® / plants
In detail: cortex of kidney / medulla tissue / liver tissue / neonatal rat cardiomyocytes / macrophages / HeLa / HEK 239 / MCF-7 / D14 SJR / D05 GSM 1A / T47D / 293 / U251 / U373 / HCT116 / GaMG / VH6-TE / garden cress
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In general: cytosolic proteins / membrane proteins / phosphorylated proteins / lipoproteins / glycoproteins / small proteins (17 kDa) / large proteins (250 kDa)
In detail: Cyclin D1 / p53 / Cleaved Casp-3 / Bax / Erk2 / RNA binding protein / b-Actin / a-Actinin-4 / phospho-ERK1/2 / total-ERK1/2 / g-Tubulin / MMP1 (active) / pro-MMP / E-Cadherin / prion protein / AMP activated kinase (AMPK)
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* The amount of DNA contamination is significantly reduced during on-column rDNase digestion. Anyhow, 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
** Kits to be used for research purposes only
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