RNA Stabilization and Protein Stabilization in Tissue Extracts with Stabilyser™ Reagent
Introduction
Maintain nucleic acids and functional proteins with Stabilyser™ reagent, the only reagent that protects the integrity of nucleic acid and active protein in one uniform lysate mixture. Now, you can stabilize complete, uniform tissue homogenates providing sample-to-sample comparability.
The convenient all-in-one Stabilyser™ formulation protects analytes at the time of lysis and provides protection from freeze/thaw cycles during long-term storage for future detection needs:
- Maintain functionally active protein and nucleic acids from the same tissue samples
- Long-term storage and protection from freeze/thaw cycles for future detection needs
- Archive tissue samples if future analyte needs change
- Compatible with Western blotting protocols, functional protein activity assays, and nucleic acid applications like qPCR
Procedure: RNA stabilization and/or Protein stabilization
The volume of Stabilyser™ Reagent to be added to the tissue varies based on tissue type and desired protein or nucleic acid concentration required.
In general, it is recommended to start optimization with 20 µL of Stabilyser™ Reagent per mg of tissue.
For best results, it is recommended to use fresh or flash frozen tissue samples prepared directly after harvest. Flash frozen tissue should remain frozen until added to the Stabilyser™ Reagent and homogenized with the mechanical homogenizer of choice.
For some applications, lysis at 4 °C and the addition of protease and/or phosphatase inhibitors might be advantageous.
- Determine tissue weight and aliquot appropriate volume of Stabilyser™ Reagent into a clean tube compatible with the volume of reagent needed and mechanical homogenizer being used during tissue disruption.
- Add fresh or flash frozen tissue into pre-aliquoted Stabilyser™ Reagent and immediately perform the mechanical homogenization of the tissue.
- Centrifuge the homogenized sample for 15 minutes at 12,000–20,000 x g to pellet debris.
- Remove clarified supernatant to a clean tube.
- For long term storage, it is recommended to store lysate at -70 °C.
Protein stabilization with Stabilyser™ reagent retains enzyme activity even after a day at room temperature.
RNA stabilization: CT values for GAPDH equivalent even after 24 hours at room temperature.