Introduction 22
P/N CLS151164 Rev. B LabChip GX Touch/GXII Touch User Manual PerkinElmer, Inc.
Principles of Operation (Continued)
For DNA, RNA, Protein, and Glycan assays, quantitating the
concentration and accurately sizing each fragment are achieved by
comparing against a sizing ladder and running internal standards or
“markers” with each sample. Internal standards of known
concentration are mixed with the sample to aid in quantitation.
The amount of sample sipped into the chip depends on pH, salt
concentration, and buffer additives. The internal standards
normalize these factors so that the software can use the ratio of the
area of the curve of the standard to the unknown peak to determine
concentration directly. The internal standards lie slightly outside the
assay range so they do not interfere with analysis.
Capillary Zone Electrophoresis (CZE) is an electrophoretic
separation technique used to evaluate the charge heterogeneity of
proteins in a sample. For Protein Charge Variant assays, analytes
are separated based on their net charges, with molecules with a
higher net charge migrating faster than those with a lower net
charge.
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