PureProteome磁力架

Comparison of elution techniques for small-scale protein purification of FLAG® tag proteins using anti-FLAG® M2 magnetic beads.

PureProteome™ Protein A and G Magnetic beads provide a rapid and reproducible means to purify immunoglobulins (IgG) using the KingFisher Duo particle processor.

比较anti-FLAG® M2磁珠的小规模FLAG®标签蛋白纯化的不同洗脱方法。

通过 PureProteome™ Protein A 和 Protein G 磁珠，可利用 KingFisher Duo 磁珠纯化仪快速、可重现地纯化免疫球蛋白 (IgG)。

Biomarkers offer important information about homeostasis, disease, response to drug treatments, and environmental stimuli. Sera are rich sources of biomarkers (biological indicator proteins, peptides, small molecules, etc.) and are easier to sample than other tissues. However, the complexity of serum and the presence of highly abundant proteins like albumin and immunoglobulin can mask less abundant species, hindering biomarker detection. PureProteome albumin magnetic beads remove more than 98% of albumin from human serum. Here, we demonstrate that PureProteome albumin magnetic beads may also be used to remove albumin from mouse, guinea pig and rat sera. Depleted samples are often dilute, and may need concentration for downstream analyses. Therefore, we present a protocol for the convenient concentration of these samples using Amicon Ultra 2 mL centrifugal filters.

Traditionally, protein purification from E. coli consists of four distinct phases: harvest, bacterial cell lysis, lysate clarification and protein purification. Bacterial lysis typically requires several time-consuming, hands-on steps, such as freeze/thaw cycles and sonication. These harsh lysis techniques may negatively impact protein quality and contribute to sample-to-sample variability. To maintain protein activity and integrity, detergent-based lysis buffers are routinely used to avoid mechanical protein extraction methods. Regardless of the lysis method used, centrifugation is traditionally required to pellet unwanted cell debris and permit recovery of the clarified lysate. The final step, purification, is frequently performed using affinity media specific for expressed epitope tags. Agarose-based media have typically been used, either as a slurry in microcentrifuge tubes or packed into gravity-driven or spin columns. While easier to manipulate, columns are greatly affected by lysate consistency and carryover of cell debris, which can lead to clogging of the column frits.