User:Paul Rothenberg/Notebook/CHEM 571 Fall 2014/2014/09/23
|Project name||Main project page|
Previous entry Next entry
SDS-PAGE of Lysozyme
We are running SDS-PAGE of the samples that Monika prepared. Here is what she did:
5 samples were prepared and stored in the fridge for electrophoresis next week.
The protocol can be found here. The proteins solutions prepared were placed in 100°C in a thermocycler for 5 minutes to denature the protein. The running buffer was diluted 10X.
20 µL of the protein ladder was loaded, along with 20 µL of samples in each well. The ladder was in well 1, BSA in 3, BSA colloid in 5, Lysozyme in 7, Lysozyme colloid in 9, and Soy Protein in 11.
The gel was run for 30 minutes at 200V. It was fixed for 30 minutes, stained for 1 hour, and destained for 15 minutes. The gel was further rinsed by Dr. Fox.
Redo of Bradford Assay
The data was not conclusive for the Bradford assay, so we are re-running it in larger quantities with serial dilutions. 1L of 0.9% saline was prepared, and 0.50350 g of lysozyme was dissolved. Serial dilutions were prepared with the 50 mM Tris and NaCl buffer by a factor of 2 each time.