NucleoSpin RNA/Protein

NucleoSpin^® RNA/Protein
Total RNA and protein isolation

Features

Parallel isolation of RNA and protein from unsplitted samples

• 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.
• Parallel purification of genomic DNA possible by using the NucleoSpin RNA/DNA
   Buffer Set



Technology
Silica-membrane technology

Format
Mini spin columns

Sample material
< 5 x 10⁶ cultured cells
< 30 mg human/animal tissue
< 100 mg plant tissue

Total RNA

Total protein

Fragment size
200 b - 20 kb

15 - 300 kDa

Typical yield
< 70 µg

< 1200 µg

A_260/280
1.9-2.1

Does not apply

Elution volume (RNA)
Resolubilization volume (protein)
40 - 100 µl

10 - 100 µl

Preparation time
30 min/6 preps

35 min/6 preps

Binding capacity
200 µg

Does not apply

Applications**

• 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

Starting materials successfully used Proteins successfully analyzed

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

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)

* 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