Protocol: Protein Extraction with Ossiform P3D Scaffolds
This protocol explains how to perform protein extraction with Ossiform P3D Scaffolds.
- Lysis buffer
Figure 1.: Workflow for harvesting protein from cells cultured on P3D Scaffolds.
Notes before starting and general advice on material handling
- All handling of the P3D Scaffolds products should be performed using gloves, according to the standard aseptic methods.
- The scaffolds are supplied sterile by dry heat sterilization and remains sterile until opened.
- Aspirate cell medium.
- Wash 10-20x in prewarmed PBS for 1 minute each wash. It is important to remove all trace of medium.
- Cover scaffolds in lysis buffer.
- Shake scaffolds at 300 rpms for 30 min.
- Flush scaffolds and collect lysate in Eppendorf tubes.
- Centrifuge samples at 16,000 g for 20 min at 4C.
- Place samples on ice and pipette supernatant with proteins in to clean prelabelled Eppendorf tubes.
- Samples can now be stored at -80C and continue to downstream protein applications.
The products are “For Research Use Only (RUO)” and should not be used for clinical, diagnostic, or therapeutic purposes. Ossiform and its distributor, Ilex Life Sciences LLC, make no other warranties, expressed or implied, including the implied warranty of merchantability and the implied warranty of fitness for particular purpose.
This protocol has been adapted from the protocol published by Ossiform. Ilex Life Sciences is an authorized distributor of Ossiform P3D Scaffolds research product line.