Hoefer SE250 - Manual (Page 18)

Note: The amount of protein
sample added to each well
depends on both the sensitivity
of the staining method and
the distribution of protein
among separate bands. With
Coomassie
™ Blue, it is possible
to detect 1 µg in a single band;
with the more sensitive silver
stains, it is possible to detect as
little as 10 ng.
3
To aid in loading samples, wet the well-locating decal
and apply it to the front of the glass plate so that the
appropriate edge outlines the sample wells.
Note: The side wells for standards of a preparative
comb correspond to the outer-most wells formed by
the 10-well comb.
4
Fill the sample wells and each upper buffer chamber
that will be used with running buffer. One upper
buffer chamber holds approximately 75 mL.
5
Underlay the sample into the wells using a fine-tipped
microsyringe. The width of the wells depends on the
number of wells per comb. If the comb has fewer
wells, they are wider, and require more volume to
raise the level 1 mm, as shown in the following table.
Volume of sample (µL) per 1 mm depth
no. of comb thickness (mm)
wells 0.75 1.0 1.5
5 9.5 12.7 19.1
9 5.8
10 3.6 4.8 7.2
15 2.2 2.9 4.4
18 2.9
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p10