Sample quality is imperative to generating good quality data from Biacore interaction analyses. Users should adhere to the following guidelines for all samples submitted to or used on the BIA Core Facility’s instruments.
- Samples should be highly purified (single band by gel electrophoresis).
- Samples should be checked for homogeneity by size exclusion chromatography, and any aggregates or fragments present should be removed (>95% purity by SEC is ideal).
- Sample concentration should be determined as precisely as possible and extinction coefficients and MWs should be provided.
- Sample concentration is recommended to be at least 1 mg/mL.
- The recommended amount of sample required is between 50-100 µg.
- Buffer compatibility of samples should be verified with the facility. The BIA Core Facility regularly uses HBS-EP+, HBS, and PBS as running buffers. If a specific running buffer is desired, it must be provided by the user (0.22 µm filtered and degassed).
- Cryoprotectants (such as glycerol), if present, should be removed.
- If biotinylated samples are to be used in experiments, the efficiency of biotinylation should be verified.
- Any gel and/or SEC purity profiles should be provided.
- Any ELISA data or Kd values determined by other techniques should be provided.
Arrangements for sample drop-off should be made with the Facility Manager or the appropriate BIA Core staff member. Please ensure that all sample tubes are clearly labelled, and the completed Sample Submission Form is attached to the CoreResearch@Duke Service Request.