Dna protein binding equation and percent saturation
Answers
hello mate ⚘
However I wish to state here that my DNA is fluorescently labelled at 5' terminus and 4–6 nM fluorescein end-labeled nucleic acid is needed to achieve sufficient signal- for a quantitative measurement. Since, it has been suggested in various studies that for proteins that bind to their target sequences with an apparent Kd <20–40 nM, the trace assumption associated with fitting the Hill equation is violated. In such cases, it may be necessary to fit the data to a quadratic equation as shown in the picture (Equation 3, where R is labeled nucleic acid concentration, P is the total protein concentration). Failure to do so will lead to over-estimation of the equilibrium dissociation constant and Hill coefficient. (Ryder, RNA; 2011)
I have been advised that i cannot use the binding equation Y=Bmax*X/Kd+X (where x is enzyme conc and y is percent dna bound) under my assay conditions.
I have been struggling hard to apply this equation as i cant find it in any software like prism origin by default. However when i am trying to enter my equation in the softwares its giving me error.. Could someone please help me with that.
