B/B0 Ratio Outside Acceptable Range at Curve Endpoints

Symptom

The B/B0 ratio (bound/maximum bound) for the terminal point of the standard curve is either >95% or <5%, indicating insufficient competitive displacement range.

1. 1 Standard concentration range not optimized for the competitive binding dynamics
2. 2 Highest standard concentration insufficient to achieve >95% competition
3. 3 Lowest standard concentration causing excessive competition beyond useful detection limit

1. 1 Increase standard concentration by one dilution factor (e.g., 2× or 3×) to shift curve upward if B/B0 >95%
2. 2 Decrease standard concentration by one dilution factor to shift curve downward if B/B0 <5%
3. 3 Re-establish standard curve with 7-8 points spanning 2-3 orders of magnitude around expected sample concentrations