Optimization of an HP Scanjet for quantification of protein electrophoresis gels.
An inexpensive desktop scanner, the Hewlett-Packard Scanjet IIp (HP), has been optimized for analysis of protein electrophoresis gels by comparison with a calibrated laser densitometer (Laser). Images from both densitometers were transferred to a personal computer and analyzed with QGEL software. Without correction the HP response was often in poor agreement with the Laser. However, when the HP response to Coomassie blue stained gels and x-ray films was linearized using a HP software option called Emphasis, the HP results agreed with results from the Laser. For 2D gels scanned with appropriate Emphasis applied, spot integrated density values were a constant multiple of 1.8 +/- 0.3 times the corresponding Laser value for x-ray films (CV = 17%) and 2.1 +/- 0.5 for Coomassie blue stained gels (CV = 24%). The highest error was observed for density extremes. For proteins quantified relative to standards using sodium dodecyl sulfate-slab gel electrophoresis, the HP values were within 15% of the Laser values. Data is shown concerning linearity and reproducibility of response, optical density range (about 0 to 1.8 OD units), variability of the imaging field, and resolution of the HP.
Kendrick
N C
NC
Kendrick Laboratories, Inc., Madison, Wisconsin 53713.
Johansen
J J
JJ
Lee
P R
PR
Santek
D A
DA
D001345:Autoradiography / Q000379:methods; D002250:Carbon Radioisotopes; D003720:Densitometry / Q000295:instrumentation* / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004926:Escherichia coli / Q000378:metabolism; D007202:Indicators and Reagents; D007834:Lasers; D010452:Peptide Biosynthesis; D010455:Peptides / Q000032:analysis* / Q000302:isolation & purification; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification; D012394:Rosaniline Dyes; D012710:Serum Albumin, Bovine / Q000032:analysis; D012984:Software; D013462:Sulfur Radioisotopes
D002250:Carbon Radioisotopes; D007202:Indicators and Reagents; D010455:Peptides; D011506:Proteins; D012394:Rosaniline Dyes; D013462:Sulfur Radioisotopes; D012710:Serum Albumin, Bovine; C048139:Coomassie blue