SDS-PAGE Analysis
Reagents Required
Recipes are provided in Appendix 3, page 31.
6X SDS Loading Buffer
1. Remove 10 μl of each supernatant from Step 14 (above)
to fresh tubes.
2. To these aliquots, and to the 10 μl samples retained
following steps 7 and 9 (above), add 2 μl of 6X SDS
Loading Buffer.
3. Vortex briefly and heat for 5 minutes at 90-100°C.
4. Load the samples onto a 10-12.5% SDS-polyacrylamide gel.
5. Run the gel for the appropriate length of time and stain
with Coomassie Blue to visualize the parental GST (made
in control cells carrying the parental pGEX vector) and
the fusion protein.