To synthesize AuNP using the procedure developed here:
Bakshi, Mandeep S.; Kaur, Harpreet; Khullar, Poonam, Banipal, Tarlok, S.; Kaur, Gurinder; Singh, Narpinder Journal of Physical Chemistry C 2011, 115(7), 2982-2992
In addition, to repeat the experiment performed on 08/31/11, 09/07/11, 09/27/11, 09/28/11, and 10/19/11.
To run an SDS-PAGE to determine whether or not MBP-intein fusion proteins are being expressed cells comparable to the cells frozen on 09/28/11 and to continue the experiment on 10/25/11.
- Combine 2 mL HAuCl4, 1 mL BSA, and 7 mL of water in a test tube.
- Combine 1 mL BSA and 9 mL of water in a separate test tube.
- Place test tubes in an oven at 80°C for 30 min.
- Remove test tubes from oven for 10 min.
- Repeat steps 3 and 4 for 3.5 hr.
- Empty Coomassie Brilliant Blue R250 stain from gels.
- Add destaining solution.
- Place gels on shaker for 2 hr.
- Dump out destaining solution.
- Add more destaining solution.
- Place on shaker overnight.
After 30 min in the oven, yellow fibers began to appear at the surface of the gold-containing solution. At 60 min, a greenish-gold solid began to form near the surface of the solution.
The solution containing only BSA and water remained clear throughout the experiment.
The gels indicated that the desired protein was present, but it was not pure.
Protein solutions used were from group 2: