User:Eric Ma/Notebook/MICB323 Lab Book/2009/03/10

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Pre-Lab Calculations for SDS-PAGE

 * Note: Unable to do calculations for recombinant virus. Did calculations for Wild-Type virus based on BCA assay results. Did not use BSA Assay results.

Calculations for BCA Assay using WT Virus

 * Sunny did an average of the two volumes and pipetted them in.
 * For the recombinant virus samples, we simply pipetted in 15 µL of each sample + 3 µL of 6X loading buffer.

Part 1: Determination of Protein Concentration of LPS-Stimulated Cel Lysates using Bradford Assay

 * Did this together with Sunny.

Part 2: SDS-PAGE of Insect Viral Lysates

 * Prepared two gels: one for myself, one for Sunny.
 * My gel is the gel that had lots of bubbles, and hence not enough wells to load in all the samples.
 * Sunny's gel is perfect. Therefore, Sunny will run the entire 15 samples, while I will omit all of the Recombinant virus lysates.

Comments on the trends

 * Stimulation with LPS
 * We see that the stimulation with LPS results in a peak of protein production at 24 hrs p.s. (post stimulation), before gradually decreasing at 48 hrs and 72 hrs p.s.
 * Stimulation with IFN-g
 * We see that the stimulation with IFN-g results in a peak protein production at 24 hrs p.s., gradually decreasing at 48 hrs p.s., and dramatically decreasing at 72 hrs p.s.
 * Combined stimulation with LPS and IFN-g
 * We do not see an observable trend. First, there is a decrease in protein concentration from 24 hrs p.s. to 48 hrs p.s., and then an increase at 72 hrs p.s.

Calculations
Full calculations can be found in [[Media:090310_Lab_7_Calculations.xlsx|this]] file.


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