Sample Blank BSA (2 ug/uL) BSA (1.5 ug/uL) BSA (1 ug/uL) BSA (0.75 ug/uL) BSA (0.5 ug/uL) BSA (0.25 ug/uL) BSA (0.125 ug/uL) BSA (0.025 ug/uL) Protein Extract (C) Protein Extract (T) Questions Abs 1 0.231 2. 2.666 3. 2.213 1.642 1.373 1.027 0.677 0.466 0.284 0.855 0.863 Abs 2 0.238 2.714 2.269 1.661 1.342 Protein (g/L) Protein (g) Volume of protein to load (L) 2x LB (L) 1.041 0.67 0.466 0.276 0.859 0.871 Average Abs 0.2345 Table 4.3 Pipetting scheme for a Western Blot. 2.690 2.241 ? 1.6515 The instructor will provide you with BSA Assay data (posted on Blackboard) that will allow you to determine the concentration of protein in the RIPA lysate samples prepared by the Laboratory Instructor. Use this information to: 20 1.3575 1. Determine the concentration of total protein in each of the RIPA cell lysate samples for the control (C2C12 myoblasts, C) and the treated (C2C12 myotubes, T) cells. Show all calculations and include all units where necessary. ? 1.034 ? 0.6735 0.466 Determine the amount of sample volume required to load 20g of protein for each sample using the concentrations determined in Question 1. Show all calculations and include all units where necessary. 0.28 Using MS Excel (or similar) prepare a data table (refer to Table 4.3 below for reference) that outlines the preparation of samples to be loaded into the polyacrylamide gel for electrophoretic separation. The volume of 2x Laemmli Buffer (LB) to be added is equal to the volume of the protein sample (1:1 dilution). 0.857 0.867 Myoblasts 1 (C1) Myoblasts 2 (C2) ? 20 ? ? Myotubes 1 (T1) ? 20 ? ? Myotubes 2 (T2) ? 20 ? ?