AlphaLISA Anti-M13 Acceptor Beads, 250 ug

Validation of Alpha Anti-M13 beads for detecting protein-protein interactions

The Alpha Anti-M13 Acceptor beads were validated to detect protein-protein interactions (PPI) commonly used in phage display technology. In this case study, M13 phages designed to express VHH anti-human HER2 and selected by phage display were produced in E. coli in a 2YT bacterial medium. The PPI assay was performed in a 384-well shallow plate by adding 5 µL of sample, 5 µL of biotinylated human HER2, 5 µL of Alpha Anti-M13 acceptor beads at 20 µg/mL, and 5 µL of Alpha SA-Donor beads (#6760002) at 40 µg/mL. The mixture was then incubated for 20 hours at 23°C in the dark. Detection reagents were prepared in Immunoassay buffer (AL000C/F). PPI interaction was measured by recording the Alpha signal.

* If sample titration is required, It is recommended to dilute in the sample buffer or medium to avoid matrix effect.

Validation of AlphaLISA Anti-M13 acceptor beads for detecting M13 phage expressing the target of interest

M13 phages designed to express VHH and selected by phage display were produced in E. coli in a 2YT bacterial medium. It is generally accepted that phage samples contain only a fraction of the expressing population (~0-10%). To validate VHH expression in this M13 production, a quality control assay was performed in a 384-well shallow plate by sequentially mixing 5 µL of unpurified sample (serially diluted*), 5 µL of IAB buffer*, 5 µL of Anti-M13 acceptor beads at 20 µg/mL, and 5 µL of Anti-VHH donor beads at 40 µg/mL (Cat#AL117). The mixture was then incubated for 20 hours at 23°C in the dark. Detection of the Alpha signal confirms that the sample contains M13 phages expressing the VHH.

* If sample titration is required, It is recommended to dilute in the sample buffer or medium to avoid matrix effect.

** Immunoassay buffer (AL000C/F) was used for this assay to dilute detection reagents.