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Imaging Chemiluminescent Western Blots

l ULTRA substrate had an integrated density measurement equivalent to a 216-ng band using ECL substrate, based on the equation of a standard curve generated when all four of the visible ECL bands were used as standards. This equates to an 11.5-fold increase in sensitivity observed when using SuperSignal ULTRA substrate compared to ECL reagent. These results suggest that researchers using the Eagle Eye II still video system to directly image filters from chemiluminescent Western blots may be able to increase sensitivity or decrease exposure time by using SuperSignal ULTRA substrate.

Conclusions

The Eagle Eye II still video system features a light-tight cabinet and a CCD camera that is sensitive to wavelengths of light emitted by common chemiluminescent substrates. These features make this system well suited for directly acquiring images from chemiluminescent Western blots. Western blot analyses performed using two different chemiluminescent substrates indicated that some substrates may perform better than others when used with the Eagle Eye II still video system. Consequently, researchers using the Eagle Eye II system to acquire images from chemiluminescent Westerns blots may be able to improve results by carefully choosing the chemiluminescent substrate.

Acknowledgments

The authors wish to thank Kerstien Padgett for helpful discussions concerning Western blot analysis.

REFERENCES

  1. Cayouette, M., Steege, M., and Moores, J. (1997) Strategies 10: 21-23.

  2. Gackstetter, T., et al. (1994) Strategies 7: 57-59.

  3. Mattson, D.L. (1996) Previews 1: 2-3.


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