Millipore Technical Publications |
 | AN- an1966en00 | |
Key Steps for Successful Immunodetection Using the SNAP i.d. Protein Detection System | ||
| Lit No: | an1966en00 | |
| Year: | 2008 | |
| The western blot is a powerful tool used extensively in protein research to detect and compare the relative levels of proteins without the need for their prior purification. Its widespread appeal is based on its overall simplicity, coupled with the high resolution of proteins separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) prior to membrane transfer. This versatile technique for the detection and characterization of proteins continues to develop with approaches that now enable more rapid electrophoresis, faster protein transfer, increased miniaturization, and higher specificity. Additionally, recent enhancements in chemiluminescent detection have increased significantly both the sensitivity and flexibility of western blots. While much attention has focused on improving these aspects of western blotting, the immunodetection step, up until now, has been a lengthy process typically requiring four hours or more of elapsed time to complete. The SNAP i.d. Protein Detection System has been developed to shorten the time required for immunodetection (i.e., blocking, washing and antibody incubations) to approximately 30 minutes. This is achieved without any additional reagent consumption (e.g., antigen, antibody or detection reagents) or any compromise in the results; sensitivity, background, and signal-to-noise ratios are the same or better than that achieved with traditional immunodetection techniques. The SNAP i.d. system’s unique design enables the use of small volumes of antibody for incubations and either polyvinylidene difluoride (PVDF) or nitrocellulose blotting membranes. Because it is vacuum-driven, extremely rapid blocking and washing of blots are achieved. The SNAP i.d. system is compatible with fluorescent, chemiluminescent, or chromogenic detection methods. Moreover, the sequence of steps required to process a western blot with the SNAP i.d. system is identical to those used in traditional immunodetection. Click the PDF above for the full document. | |
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