Bio-Rad Experion Protein Analysis Kits - Manual (Page 9)

Starter Kit Test 1 — Sizing and Relative Quantitation
1.1 Overview of Test 1
Microfluidic chips use much smaller volumes of samples and reagents than traditional slab
gels. As a result, microfluidic separations are much more sensitive to variability in pipetting
technique when compared to traditional electrophoresis methods. Proper pipetting skills
and tools are critical to a successful Experion
™ analysis.
In this test, you prepare a dilution series of the bovine g-globulin (BGG) protein, run an
Experion Pro260 analysis of the series, check the resulting electropherograms for proper
performance, examine protein sizing and relative quantitation results, and export the data
to determine the accuracy and reproducibility of the analysis.
Since the use of proper pipet tips can also impact assay performance, use the narrow-bore
pipet tips included with the Experion Pro260 starter kit (or similar narrow-bore tips) for
loading samples and reagents into the microfluidic chip. Narrow-bore tips are required for
successful chip loading (other tips can introduce bubbles during chip loading, leading to
problems in performing the assay or in the quality of results).
1.2 Assay Procedure
1.2.1 Set Up the Electrophoresis Station
Note: If this is the first time the Experion electrophoresis station is being used, refer to
Appendix C for instructions on how to prepare the system for use.
1. Power on the computer.
2. Power on the Experion electrophoresis station. Push the green button in the center of
the front panel. The steady green LED above the button indicates that the unit is on.
3. Launch Experion software. Check the screen to confirm that the instrument and
computer are communicating properly.
When communication has been established:
•
A green dot with the last four digits of the instrument serial number appears in
the lower right corner of the main software screen
• The electrophoresis station icon appears in the upper left corner
When there is no connection, these indicators are absent and a “disconnected”
message appears next to the Start Run button
in the upper left corner of the
screen. In addition, a grayed-out instrument icon appears in the upper left corner of the
software screen.
1.2.2 Equilibrate the Kit Reagents
1. Set a heating block or water bath to 95–100°C. You will use this heating block to
denature the samples and the Pro260 ladder later in the protocol.
2. Remove the kit components from storage.
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