Direct detection of histidine tagged biomolecules on electrophoretic gel
OwnerA. Krishna Malliainventor
Life of this patent
Estimating the term…
Loading patent…
OwnerA. Krishna Malliainventor
Estimating the term…
A method is disclosed for the detection of a His-tagged target biomolecule while said molecule remains in the gel in which it has been separated from other constituents by electrophoresis. The method involves, while the separated His-tagged biomolecule remains in the gel, forming a chelate between the His-tagged molecule, a lanthanide metal ion which exhibits fluorescence, and a sensitizer which enhances the fluorescence of the ion. The gel is then exposed to a u.v. light source to thereby generate a fluorogenic signal indicating the presence of the chelate and, in turn, the His-tagged biomolecule. The fluorogenic signal can then be detected either by direct visualization and/or by other means, such as photographically, e.g., film or charged coupled device (CCD) imaging.
A method for the direct detection of a histidine tagged biomolecule on an electrophoretic gel comprising (1) separating the biomolecule from other substances by electrophoresis on a gel, (2) while said molecule remains so separated, forming a chelate between said molecule, a lanthanide metal ion which exhibits fluorescence, and a sensitizer which enhances the fluorescence of the ion, and (3) exposing the gel containing said chelate to a u.v. light source thereby generating a fluorescent signal indicative of the presence of the chelate and, in turn, the histidine tagged biomolecule, and (3) detecting the signal so generated.
A kit for the direct detection of histidine tagged biomolecules on an electrophoretic gel, said kit containing two separately packaged components and a written instruction sheet, one of said components being an aqueous solution of a lanthanide metal ion which exhibits fluorescence and the other of said components being an aqueous solution of a sensitizer which enhances the fluorescence of the ion.
Dependencies are inferred from the claim text.