Figure 1. RNA and Protein from Tissues Stored in RNAlater® for >1 Year. RNA and protein were isolated with the PARIS™ Kit from ~50 mg of mouse brain or kidney stored for 13 months at -20°C in RNAlater. Purified RNA (1 µg) was analyzed on a denaturing agarose gel and subsequently transferred, blotted, and hybridized using the NorthernMax®-Gly Kit (Ambion). A portion of the protein fraction (10 µg) was used for Western blot analysis with antibodies specific for GAPDH or Hur proteins.
Figure 2. RNA and Protein Isolation from Stabilized Samples. Isolation from ~30 mg of mouse liver, brain or kidney 5 days after collection. Samples were either kept frozen at -80ºC, or stored in RNAlater® at 4ºC.
As shown in Figure 1, no obvious RNA degradation was observed by denaturing agarose gel analysis or by Northern blot hybridized with probes specific for U1 snRNA, or ß-actin and GAPDH mRNAs. Analysis of the protein fraction by Western blot showed that the recovered proteins were full-length. (Note: The multiple bands observed in the Hur blot correspond to brain-specific higher molecular weight isoforms of the Hur protein.) Figure 2 further confirms that protein yield and quality are not affected by RNAlater treatment. These results demonstrate that RNAlater is an efficient sample collection/stabilization reagent for protecting RNA, and that RNAlater-treated samples can be used for routine protein analysis such as Western blotting.
It is especially useful for the following applications:
- Archiving samples for future analysis
- Collecting samples at different time points without having to process them immediately
- Shipping samples without dry ice
- Collecting samples in the field