Bio-Rad Experion Protein Analysis Kits - Manual (Page 56)

of protein concentration, and the sample protein concentration is then determined from this
calibration curve and reported in the the Calib . Conc . column of the Results tab. Absolute
quantitation methods are often more accurate than relative concentration methods
How Experion Pro260 Analysis Differs from SDS-PAGE
Though the Experion Pro260 analysis kit and SDS-PAGE both use a gel-based matrix and voltage
difference to separate proteins according their size, there are a number of fundamental differences
between the two methods that may affect how specific proteins are separated and how data are
interpreted (see Table A1 below or refer to bulletins 5299, 5423, and 5453).
Table A.1. Comparison of Experion Pro260 analysis and SDS-PAGE.
Aspect SDS-PAGE Experion Pro260 Analysis
Sieving polymer (matrix) Formulation can be changed to affect resolution Fixed formulation
Crosslinked polymer Polymer (not crosslinked)
Protein stain Multiple available Single, proprietary stain
Binds proteins Binds LDS micelle-protein complexes
Detection Separate staining step, proteins stained after
separation Integrated separation step; proteins stained
during separation
Separate destaining step Integrated destaining; proteins destained after
separation
Requires photography or other imaging
equipment for documentation, analysis Laser excites the stain, detection using a
photodiode, both incorporated
Types of separation Denaturing and native; reducing and
nonreducing conditions Denaturing; reducing and nonreducing conditions
Downstream applications Separated proteins available for excision,
electroelution, blotting applications Proteins not available for further analysis
Reagent and sample usage Moderate to extensive (ml to L) Minimal (µl)
Waste generation Moderate to extensive (ml to L) Minimal (µl)
Overall analysis time Hours (depends on gel size, staining method,
and methods of documentation and analysis) 30 min/chip (10 samples per chip)
Data analysis Manual; requires additional equipment and
software Automated (with flexibility of manual adjustments)
52 Technical Support: 1-800-4BIORAD • 1-800-424-6723 • www.bio-rad.com
Experion Automated Electrophoresis System