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A Better Way to Isolate RNA AND Protein From the Same Sample

E permits transition of previously frozen samples to a state compatible with handling at room temperature and standard homogenization methods (see the article"Stop Grinding Frozen Tissue" in this issue). As shown in Figure 3, the PARIS procedure is compatible with both of these tissue collection/RNA stabilization reagents for both cells and tissue. After homogenization, protein samples are readily available for applications that do not require native functional protein, such as Western blot analysis.

Figure 2. 2D Analysis of Proteins Isolated Using the PARIS Kit. Protein samples were resolved using a pH 4-7 IPG gel followed by 8-16% SDS-PAGE. Top: Total protein (~25 g) from HeLa cells stained with SYPRO Ruby. Bottom: Total protein (~125 g) from mouse brain stained with Coomassie Blue.

Figure 3. RNA and Protein Isolation from Stabilized Samples. (A) Isolation from ~30 mg of mouse liver, brain or kidney 5 days after collection. Samples were either kept frozen at -80C, stored in RNAlater at 4C, or snap frozen and then stored in RNAlater-ICE at -20C. (B) Isolation from 106 HeLa cells freshly harvested, kept 2 days in RNAlater at RT, or in RNAlater-ICE at -20C.

Figure 4. Analysis of the RNAi Effect. (A) 5 x 104 HeLa cells were transfected in a 24 well plate with the indicated plasmids. 48 hours after transfection, total RNA and protein were isolated with the PARIS Kit and analyzed by denaturing agarose gel electrophoresis, Northern blot, or Western blot. SCR represents a scrambl
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