DK79 Posted October 30, 2017 Posted October 30, 2017 I have performed a binding experiment using streptavidin agarose beads and a biotynilated molecule ranging from high to low concentration. The molecule exhibits close to 100% capture at low concentration as is expected then gradually drops off. The drop -off is more extreme at higher concentrations. If the beads become saturated past a certain concentration, should the drop off be at a steady rate? Why is there seemingly a slow drop-ff initially then a fast one (it should be noted that the % captured is measured by fluorescence) Binding curve is below. The beads are Thermo Scientific 7.5% Agarose slurry. Thanks!
CharonY Posted October 30, 2017 Posted October 30, 2017 It is not at all unusual, the laws of mass actions predict non-linearity. Read up on "fractional occupancy" in the context of Clark's theory.
DK79 Posted October 30, 2017 Author Posted October 30, 2017 i will read up on it. I know i have seem this type of curve in the past but have not been able to explain its nature. Thank you!!
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