![]() The results indicated that the CVs for a fixed number of beads were evidently smaller than the CVs for varying bead numbers, especially in the range of 1 pg/mL to 100 pg/mL, where the CVs for 100 beads were less than 10%. The other three calibration curves used a fixed number of beads, (100, 49, and 25). One calibration curve was constructed by using varying bead numbers (50–160) in the process. Then, four calibration curves were established. First, the bead screening chips were developed to extract exactly 100, 49, and 25 magnetic beads with diameters of less than 6 μm. Here, we investigated whether a fixed number of beads in this bead-based digital microfluidic immunoassay could provide more stable results. However, the bead number in each measurement was not the same, causing an unstable coefficient of variation (CV) in the calibration curve. ![]() Previously, our group developed a magnetic-beads-based digital microfluidic immunoassay with a bead number of around 100, requiring less than 1 μL of sample volume to achieve a pg/mL level limit of detection (LOD). ![]() IP performance is equivalent to or better than Protein A magnetic beads from other suppliers.In a biomedical diagnosis with a limited sample volume and low concentration, droplet-based microfluidics, also called digital microfluidics, becomes a very attractive approach. The beads are washed and then the target is eluted with low pH elution buffer. Antibody or antigen/antibody complex (IP) is first captured on the magnetic beads. The protocol for the Pierce Protein A beads has been optimized to allow for high recovery and high purity of the isolated antibody or antigen. Protein A can bind to antibodies from many different species, including mouse, human, rabbit, pig, dog, and cat. Thermo Scientific Pierce Protein A Magnetic Beads are used for purifying antibody from serum, cell culture supernatant or ascites, as well as for IP/Co-IP of antigens from cell or tissue extracts. The Pierce Protein A Magnetic Beads can be used manually with a magnetic stand, as well as with automated platforms, such as the Thermo Scientific KingFisher Instruments, for high-throughput workflows. Using our crosslinker chemistry, you can immobilize an antibody onto the magnetic particle and prevent IgG contamination in IP or Co-IP experiments. Immunoprecipitation assays performed with protein A coated beads result in high yield of target antigen with very low background. Recombinant protein A, covalently attached to a blocked magnetic bead surface, can bind to antibodies from many different species, enabling purification of antibodies from crude extracts. Versatile – beads are compatible with manual and automated workflows (e.g., Thermo Scientific KingFisher Instruments). ![]() Consistent – magnetic beads eliminate resin loss and provide for more efficient separation than traditional IP methods that use only centrifugation.Low non-specific binding – stable, pre-blocked beads provide highly purified product (e.g., antigen eluted in IP with antibody is devoid of contaminating protein from complex cell lysate).High efficiency – equivalent or higher yield of IP target antigens than magnetic beads from other suppliers.Thermo Scientific Pierce Protein A Magnetic Beads are high-capacity and high-throughput affinity particles for antibody purification and immunoprecipitation methods using manual or robotic magnetic separators. ![]()
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