Exercise 4.5 - Documentation
Figure 4.8 Silver stained protein gel
- Polaroid camera (Fotodyne Foto/Phoresis I or equivalent)
- 35 mm camera equipped with macro lens
- Stained gel
- Photograph the gels.
- Use the photographs or negatives to measure the distance
from the point of protein application (or for two gel
systems, the line separating the stacking and separating
gels) to the final location of the tracking dye near the
bottom of the gel.
- Measure the distance from the point of origin to the
center of each band appearing on the gel.
- Divide each of the values obtained in Step 3 by that
obtained in Step 2 to obtain the relative mobility (the Rf
value) for each band.
- Using either the graph of Rf values and molecular
weights from Exercise 4.2, or Figure 4.8, compute the
molecular weights of each band.
Scan the negative with a densitometer and compute Rf
values based on the distances from the point of origin to
the peak tracing for each protein band. Integration of the
area of each peak will yield quantitative data as well as
the molecular weight.
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Cell Biology Laboratory Manual
Dr. William H. Heidcamp, Biology Department, Gustavus Adolphus College,
St. Peter, MN 56082 -- email@example.com