TY  - JOUR
AB  - Comparative two-dimensional fluorescence gel electrophoresis (CoFGE) was developed to enable reproducibility of coordinate assignment for protein spots. To that end, it uses an internal grid formed by a set of pure proteins. The composition of the reference compounds is known, so that, in addition to the use of the marker nodes as anchors for gel warping, their fluorescence signal can serve to estimate the amount of protein present in an analyte gel spot of interest. Here, experimental data are provided to that effect. The reference mixture was distributed in five concentrations across the gel, thus allowing the generation of calibration curves for each marker protein. Quantification was evaluated using replicate separations of Escherichia coli lysate. Expectedly, the method worked best for well-defined proteins spots and could then be performed at less than 20% error. A suitable protein mixture (8-97 kDa) has been composed for this purpose. An abundance of reference slots (40) allows the user to alternate the concentration of the protein marker mix on the gel so that areas of gel distortion can be excluded from the calculation.
AU  - Nippes, Alina
AU  - Ackermann, Doreen
AU  - König, Simone
DA  - 2019-05
KW  - Vergleichende Fluoreszenz Gelelektrophorese
KW  - CoFGE
KW  - Proteinkoordinaten
KW  - Referenzgitter
KW  - interner Standard
KW  - Quantifizierung
KW  - comparative fluorescence gel electrophoresis
KW  - protein coordinates
KW  - reference grid
KW  - internal standard
KW  - quantification
LA  - eng
IS  - Mercator Journal of Biomolecular Analysis
M2  - 14
PY  - 2019-05
SP  - 14-21
T2  - Mercator Journal of Biomolecular Analysis
TI  - Quantification with Comparative Fluorescence Gel Electrophoresis (CoFGE)
UR  - https://nbn-resolving.org/urn:nbn:de:hbz:6-74199681152
Y2  - 2024-11-22T00:24:13
ER  -