Millipore Technical Publications |
 | AN- an1965en00 | |
Triple Well Blot Holder as a Tool for 30-Minute Western Blotting Optimization with the SNAP i.d. Protein Detection System | ||
| Lit No: | an1965en00 | |
| Year: | 2008 | |
| Optimization of antibody concentration is a key requirement for successful immunodetection with both standard methodologies as well as with the SNAP i.d. system. Because of the wide range of antibody avidities/affinities, all antibody manufacturers recommend optimization whenever a new antibody is first used in an application. Prior to the introduction of SNAP i.d. system, this has required a significant effort on the part of the user and typically involved a series of time- and labor-intensive trial-and-error experiments. The goal of this effort is to ensure that immunodetection is linear over a defined experimental range as well as to enable the routine production of publication quality western blots with little or no non-specific background signals. If a blot is incubated with sub-optimal concentrations of antibody, it will result in poor or no detection of the protein(s) of interest, or non-linear signals. If, on the other hand, a blot is incubated with a too high a concentration of antibody, the resulting background could make it impossible to discern specific from nonspecific signals. Antibody optimizations can now be accomplished in a fraction of the time using the SNAP i.d. system’s triple well blot holder and its 30-minute processing time to evaluate up to six different antibody concentrations in parallel. Click the PDF above for the full document. | |
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