ThermoFisher Scientific E-PAGE Gels, Техническое руководство

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Additional Staining Protocols,
Continued
Phosphoprotein
Staining Protocol,
Continued
5. Destain the gel with Pro-Q ® Diamond Phosphoprotein Gel destaining
solution twice for 30 minutes each time at room temperature with gentle
shaking. You may wash the gel overnight to improve sensitivity and obtain
clear background.
6. Place the gel on a UV transilluminator equipped with a standard camera
and select an appropriate filter on the camera (stain emission maxima is
555–580 nm).
You may also use a laser-based scanner with a laser line that falls within the
excitation maxima of the stain (580 nm).
7. Image the gel with a suitable camera with the appropriate filters using a
3–4 second exposure.
Results
Results obtained using an E-PAGE ™ 48 8% Gel stained with Pro-Q ® Diamond
Phosphoprotein Gel Stain are shown below.
The gel contains the following samples: (lanes not indicated are blank)
Lane Sample
1 SeeBlue ® Plus2 Pre-stained Standard (5 µL)
2, 3, 4, 5 1, 2.5, 5, and 10 µL, respectively, of BenchMark
™
Fluorescent Protein Standard
6, 7, 8, 9, 10
E. coli
lysate containing His-tagged urate oxidase protein,
35 kDa (0.05, 0.1, 0.25, 0.5, and 1 µg)
11, 12, 13, 14, 15 Negative control (
E. coli
lysate with non-His-tagged urate
oxidase 0.05, 0.1, 0.25, 0.5, and 1 µg)
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15