User:Zachary I. Mendel/Notebook/Zacks Notebook/2013/09/25
|Project name|| Main project page|
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To run SDS-PAGE to observe amount of hemoglobin remaining after yesterday's digestion. This should be a complimentary measurement to our UV-Vis analysis from yesterday.
We will be using a Bio-Rad Mini Protean system with pre-cast Mini Protean TGX gels. The manual for this system can be found here We will be running SDS-PAGE followed by gel development with Coomassie Blue staining.
Below is a short description of how we will proceed. Please refer to the manual for more detailed instructions
Table 1: Samples in Wells
|5||0.5 hrs Pepstatin|
|6||1 hr Pepsin|
|7||1 hr Pepstatin|
|9||1.5 hrs Pepstatin|
Absorbance Spectrum of Pepsin with Hemoglobin
Corrected Absorbance Spectra of 20μM Pepstatin with Hemoglobin
For our gel we placed it in 10 mls fixing solution. We then dumped out the fixing solution and stained it with 10mls of a staining solution. This stayed on for 1 hour