Western blotting is a powerful complex and sensitive technique for quantifying and detecting proteins. The results can be affected by several factors, that also include user error. Here are a few ways that user error can contribute to variability in western blots: Sample preparation: Western blotting requires careful preparation of the sample, including lysis of the [...]
There are a number of studies that have compared the accuracy and results of using the BlotCyclerTM vs. manual methods for Western blotting. Overall, the BlotCyclerTM is a valuable tool for researchers who need to perform Western blots on a regular basis. It is more accurate, reproducible, and sensitive than manual Western blotting, One study, [...]
BlotcyclerTM system’s western blot processing is highly reproducible To determine if the automated western blot processing results in highly reproducible signal, five identical blots were subjected to the analysis by BlotCyclerTM using the same antibodies as described in Fig. 2. The results on Fig. 3 demonstrated excellent reproducibility as seen visually (Fig. 3A). To confirm this quantitatively, [...]
The BlotCycler is an automated Western blot processor that has a number of benefits for those labs that perform Western blots on a regular basis. It can process multiple blots simultaneously. A typical Western blot can take several hours to complete, even when done manually. The BlotCycler can process up to 12 blots at the same [...]
Common Western blotting mistakes that can lead to inaccurate results: Using the wrong antibody. The antibody is the most important reagent in a Western blot, so it is essential to use the right antibody for the target protein. If the antibody is not specific for the target protein, it will bind to other proteins in the [...]
When processing Western Blots there is substantial evidence that primary antibody and all washing steps should be done at 4 degrees Celsius. Based on our work with both the BlotCycler Touch and Mini we have observed several benefits to running the secondary antibody at 4 degrees during the western blot processing steps. Reduced non-specific binding: Lowering [...]
The BlotCycler is an automated Western blot processing system that can save time, improve results by automating the following steps: Shaking: The BlotCycler gently shakes the blots during blocking, primary/secondary and washing steps to ensure that the reagents are evenly distributed. This helps to reduce background and improve signal-to-noise ratio. Blocking: The BlotCycler applies the [...]
The Precision Biosystems BlotCycler is an automated Western blot processor that can significantly improve the reproducibility and efficiency of Western blot experiments. It automates all the steps after protein transfer to a membrane, including blocking, incubation of primary and secondary antibodies, and washing. This eliminates the need for manual pipetting and reduces the risk of [...]
When selecting a blocking buffer for Western blotting, consider the following factors: Using a blocking buffer from the same species is important. The objective is to help prevent the primary antibody from binding to the blocking agent instead of the protein of interest. The detection method being used should be compatible with the blocking buffer. Problems can [...]
Secondary antibodies are typically polyclonal antibodies which means that they are made up of a mixture of different antibodies that recognize different epitopes on the target protein. This can therefore lead to non-specific binding to various proteins during Western blotting. Some of these antibodies may also recognize epitopes on other proteins, besides your protein of [...]
