Using the equation of the standard curve, the molecular weight of the thickest bands in lane 4 were calculated as 27.18 kD and 16.7 kD for bands a and b respectively.
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Data Analysis
Column purification samples
The column purification samples 1, 2, 3 and 4 were ran in a gel using 10 µL or 15 µL and the results were as shown in Figure 1 below. There were two darker bands in the tube #3.
Figure 1: Å gel results after staining. Two major bands (a and b) were observed in tube 3 both in 10 µL and 15 µL samples. Lane 1 – protein standards, Lane 2 – Tube 1 (15 µL), Lane 3 – tube 2 (15 µL), Lane 4 – tube 3 (15 µL), Lane 5 – cell lysate, Lane 6– protein standards, Lane 7 – Tube 1 (15 µL), Lane 8 – tube 2 (15 µL), Lane 9 – tube 3 (15 µL), Lane 10 – cell lysate
A standard curve of the protein standards was created by plotting log of molecular weight against the distance moved from the wells (Figure 3).
Figure 2: Standard Curve created by plotting log of molecular weight against distance moved by the standard proteins from the wells
Using the equation of the standard curve, the molecular weight of the thickest bands in lane 4 were calculated as 27.18 kD and 16.7 kD for bands a and b respectively.
Sample calculation using band a, which had moved 38 mm,
Ara/Amp Samples
Ara/amp samples were loaded into the wells of a NUPAGE gel and ran. The results were as shown in Figure 3 below.
Figure 3: NUPAGE gel results after staining. Two major bands (a and b) were observed in lane 4 and 8 and a single band in 5 and 10. Lane 1 – protein standards, Lane 2 – amp – heat, Lane 3 – amp – no heat, Lane 4 – ara/amp – heat, Lane 5 – ara/amp – no heat, Lane 6– protein standards, Lane 7 – amp – heat, Lane 8 – amp – no heat, Lane 9 – ara/amp – heat, Lane 10 – ara/amp – no heat
Using the equation of the standard curve, the molecular weight of the thickest bands in lane 4 and 5 were calculated as 27.18 kD, 17.7 kD, and 27.18 kD for bands a, b and c respectively
2.397940008672038 2.176091259055681 2.0 1.8750612633917 1.69897000433602 1.568201724066995 1.397940008672038 1.301029995663981 1.176091259055681 1.0 4.0 9.0 17.0 19.0 27.0 33.0 39.0 43.0 49.0 55.0
Molecular Weight ( Log kDa)
Distance (mm)