2022-10-21
Nickel Affinity Purification of Nanobodies
Equilibration
The packed nickel resin was washed with a buffer to remove the storage solution.
Sample Loading
A small portion of the periplasmic extract (PE) was saved for analysis, and the rest was loaded into the column, allowing it to drain naturally. A fraction of the flow-through was collected for analysis.
Washing
The column was washed to remove non-specifically bound proteins, and the last portion of the wash was saved for further analysis.
Elution
The bound nanobodies were eluted using an imidazole solution. The eluates were collected in small fractions for analysis.
Protein Measurement
The protein concentration of the eluates was measured using a spectrophotometer, and the fraction with the highest concentration was kept.
Storage
The protein fraction was stored at low temperature for further analysis using SDS-PAGE.
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