A new method for the targeted analysis of proteins is presented that combines the specificity of immunoprecipitation with the sensitivity of protein detection on microchips using the High Sensitivity Protein 250 assay for the Agilent 2100 Bioanalyzer. As an alternative to Western blotting, this method is valuable for researchers engaged, for example, in protein expression and purification in pharma, biotech or academic labs. Advantages of this new method in comparison to Western blotting are: higher specificity and sensitivity, less manual steps and direct availability of quantitative data, 3 hours versus 1 day analysis time, 10x less primary and no secondary antibody needed, environmentally friendly process.
The document presents a new method for the targeted analysis of proteins that combines the specificity of immunoprecipitation (IP) with the sensitivity of protein detection using the High Sensitivity Protein 250 assay for the Agilent 2100 Bioanalyzer. This approach serves as an alternative to Western blotting, offering advantages such as higher reliability, better accuracy and precision due to fewer manual steps, increased productivity with a total analysis time of about 3 hours, and lower reagent consumption.
| Total analysis time | about 3 h |
|---|---|
| Detection limit (LOD) | 100 pg/pL |
The total time demand for the IP/HSP250 method is about 3 hours.
The total time demand for the IP/HSP250 method is about 3 h.
The limit of detection was determined to 0.001 % or 100 pg/pL PTEN.