| abstract |
A method for quantifying a DNA binding protein in a biological sample. Thenmethod may be used in the absence of radioisotopes, and at a sensitivity greater thannthe commonly used electrophoretic mobility shift assay (EMSA) method previouslyndiscussed. In particular, the method includes the steps of: a) treating the sample tonobtain a liquid to be tested for the DNA binding protein, b) incubating the liquidncontaining the DNA binding protein with a DNA, containing a binding sequence tonwhich DNA binding protein can specifically bind so that DNA binding proteinnspecifically binds to the DNA, c) separating the DNA with bound DNA bindingnprotein from other DNA and proteins in the liquid, and e) quantifying the DNAnbinding protein without interference from other proteins of similar molecular weightnin the absence of the use of a radioisotope, and at a sensitivity greater than EMSA.nThe amount of DNA binding protein may be quantified in several ways, e.g. directlynby immunoreaction to the DNA binding protein or indirectly by amplification of DNAnspecifically bound to the binding protein followed by quantification of the amplifiednDNA. In a further embodiment of the invention, the total protein in the sample, havingnat least one specific immunoreactive site in common with the DNA binding protein, isnquantified and compared with the quantity of binding protein to give an indication ofnrelative proportions of bound and unbound protein in the total protein. |