The Ambion™ PARIS™ System is used to isolate RNA and native proteins from the same experimental sample. The kit also allows isolation of nuclear and cytoplasmic fractions prior to RNA and/or protein isolation. The kit contains sufficient reagents for 50 purifications each from 1 to 75 mg of tissue or 100 to 10⁷ cells.

• Fast, simple procedure
• Isolation of nuclear and cytoplasmic RNA and proteins from cultured cells
• Isolation of total RNA and proteins from cultured cells or tissues
• No phenol required Extraction or alcohol precipitation
• Reduces time, cost and variability between independent experimental samples

Isolating high-quality RNA is the first step in a variety of gene expression analyses. Often, complementation studies are also required at the protein level. Typically, these analyzes are performed using identical experimental sample aliquots. However, when working with rare, difficult to obtain, or very small samples, it is sometimes impossible to isolate RNA and native proteins independently. In studies involving large numbers of samples, expensive reagents, or inherent variability (such as cell transfection), adding independent experimental samples is not only costly and time-consuming, but can also lead to inconsistent results. To address these issues, we developed a protein and RNA isolation system (PARIS™ System). It can simultaneously isolate RNA and protein from the same experimental sample (see figure). Using the PARIS™ System, RNA and proteins can be isolated simultaneously from whole cell lysates. Alternatively, RNA and protein can be isolated from separate nuclear and cytoplasmic fractions. Total cell lysates are prepared by first mixing tissue or cultured cells in cold cell disruption buffer. Proteins and RNA can be purified directly from this lysate due to rapid homogenization both on ice and in the presence of detergent. For RNA isolation, a portion of the total cell lysate can be immediately mixed with an equal volume of lysis/binding solution. Total RNA was then purified from the mixture using an RNA-binding glass fiber filter column. After three quick wash steps, high-quality RNA is eluted in a concentrated form. The entire procedure can be completed in less than 20 minutes. Note: This kit is not recommended for use in tissues with high ribonuclease content (such as pancreas).

It is compatible with most downstream applications
RNA isolated from total, nuclear or cytoplasmic fractions using the PARIS™ procedure can be used in a variety of downstream applications, including blot hybridization,< i>In vitroTranslation, cDNA synthesis and RT-PCR. DNase I treatment of RNA used in RT-PCR experiments (see Auxiliary Products) is recommended. Protein fractions are ready for use in most common applications, including functional assays, immunoprecipitation, Western blotting, or 2D gel electrophoresis (as shown).

Auxiliary products:
TURBO™ DNA-free™ or DNA-free-free™ DNase treatment and removal reagents (SKU #AM1907 and AM1906 respectively) are ideal for rapid removal of trace amounts of DNA from total and nuclear RNA samples without the need for phenol extraction or alcohol precipitation. The cytoplasmic RNA fraction contains virtually no DNA contamination.