NucleoSpin® RNA/Protein	Total RNA and protein isolation
Features
Parallel isolation of RNA and protein from undivided samples
• Reliable interpretation of RNA and protein extracted from the same sample    → direct correlation – no splitting of samples required • High RNA yield and integrity • High protein yield independent of protein size, localization, modification, etc. • Easy protein quantification using the MACHEREY-NAGEL Protein Quantification Assay • Complete mini kit with NucleoSpin® Filters (shredders) and recombinant DNase for    DNA-free RNA of high quality

  

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	15–300 kDa
Typical yield	< 70 µg	< 1200 µg
A260/A280	1.9–2.1	–
Typical RIN (RNA integrity number)	> 9	–
Elution volume (RNA) Resolubilization volume (protein)	40–120 µL	10–100 µL
Preparation time	30 min/6 preps	35 min/6 preps
Binding capacity	200 µg	–
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 / β-Actin / α-Actinin-4 / phospho-ERK1/2 / total-ERK1/2 / γ-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