Assignment 2 .docx - Experiment 4 The Use of Western Blot...

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Experiment 4: The Use of Western Blot in Detection of -galactosidase Protein Moksha Patel Madison Dr, Hausner BLG 888 Apr 7, 2020
1 Overall Objective The objective of this experiment is to detect the presence of a protein of interest, particularly the -galactosidase, from one of the two strains of Escherichia coli , the EMG 26 K- 12 lac - (i + z - y + ) and the EMG 9 K-12 lac - (i - z + y + ) strain, by using Western Blotting techniques and electrophoresis.
2 Results Table 1. Distance travelled by protein markers of Tris-Glycine 4-20% during electrophoresis The Protein marker of Tris-Glycine 4-20% produced 12 bands. The bands were used to calculate the relative distance travelled in an SDS-polyacrylamide gel electrophoresis. The distance travelled by each band was calculated using a ruler. The molecular weight of the protein was given, the log of the molecular weight was calculated along with the Rf values. Molecular Weight (kDa) Log Molecular Weight Distance Travelled (cm) Rf value 245 2.39 0.0 0 180 2.26 0.2 0.025 135 2.13 0.5 0.064 100 2.00 1.0 0.13 75 1.88 1.5 0.19 63 1.80 2.1 0.27 48 1.68 3.0 0.38 35 1.54 4.0 0.51 25 1.40 5.3 0.68 20 1.30 5.9 0.76 17 1.23 7.0 0.90 11 1.04 7.8 1.0
3 0 0.2 0.4 0.6 0.8 1 1.2 0 0.5 1 1.5 2 2.5 3 f(x) = − 1.19 x + 2.21 R² = 0.96 Rf (cm) Log Molecular Weight

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