Bio-Rad MicroRotofor™ Cell - Manual (Page 6)

b. The cathode assembly (black) is attached to the focusing chamber
end containing the cathode membrane (black casing).
4. Align one row of focusing chamber ports with the vents on the electrode
assemblies. These are the sample loading ports. (Figure 3)
Fig. 3. Alignment of the sample loading ports with the vents on the anode (left) and
cathode (right) assemblies.
4.3 Prepare and Load the Protein Sample
Prepare the protein sample
The protein sample included in the starter kit contains three naturally colored
proteins in deionized water, Phycocyanin (pI range 4.5–5.5), Hemoglobin
(pI range 6.0–7.5), and Cytochrome c (pI range 8.0–9.0). Each protein is
present at a concentration of 2 mg/ml for a total protein concentration of
6 mg/ml.
Note: Vortex sample vial to homogenize material.
• To 100 µl of protein sample (600 µg total protein), add 150 µl Bio-lytes
pH 3-10, and 2.75 ml deionized water for a total volume of 3.0 ml.
Load the Protein Sample
The focusing chamber features two rows of ports. The row that is aligned
with the vents on the electrode assemblies will be used to load the sample
(loading ports). The row on the opposite side of the focusing chamber will be
used for harvesting the fractions (harvesting ports). The harvesting ports
must be sealed with tape before the sample is loaded. After the sample is
loaded, the loading ports will also be sealed with a piece of tape.
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