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Imaging Chemiluminescent Western Blots

Use the Eagle Eye II still video system to directly acquire images from chemiluminescent blots

Michelle Cayouette Jane Moores
Stratagene

The Eagle Eye II still video system is well suited for direct imaging of chemiluminescent Western blots. The light-tight cabinet is ideal for the longer exposure times necessary to capture chemiluminescent signals. Also, the systems charge-coupled device (CCD) camera efficiently captures those wavelengths of light emitted by common chemiluminescent substrates. A comparison of chemiluminescent substrates from two manufacturers showed greater than a 10-fold difference in signal intensity. This difference indicates that by carefully choosing the appropriate chemiluminescent substrate for their system, researchers using the Eagle Eye II still video system to image chemiluminescent Western blots may be able to enhance their results.

The Eagle Eye II still video system is a powerful digital documentation and analysis tool for scientific researchers. The system combines quick and easy image acquisition with software processing functions, such as molecular weight determinations, densitometry calculations, and colony or plaque counting.1 Because of its light-tight cabinet and optimized camera sensitivity, the system also efficiently captures images directly from chemiluminescent experiments.2 Although the Eagle Eye II still video system can capture light emitted from the wide variety of substrates typically used in chemiluminescent Southern, Northern and Western blotting techniques, we have found that the substrate choice can have an impact on the quality of the resulting images.

Chemiluminescent Western Blots: Substrate Comparison

Chemiluminescent substrates were compared using the Eagle Eye II still video system to capture an image following Western blot analysis. Dilutions o
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