Quantitative fluorescent protein standards

摘要

The present invention provides dual labeled protein standards useful for the simultaneous determination of the molecular weight of a subject protein as well as the relative mass (i.e., amount) of the subject protein present in an electrophoresis lane. The invention is also directed to methods suitable for the preparation of such dual labeled protein standards and to methods of using such dual labeled proteins to simultaneously determine the molecular weight and the relative amount of a subject protein. Further embodiments are directed to the use dual labeled protein standards to make a more accurate determination of the amount of a protein present in an electrophoresis lane. Yet further embodiments are directed to kits containing the presently described dual protein standards. Dual labeled protein standards made and used in accordance with the embodiments set forth herein may be used to simultaneously determine the molecular weight and the relative amount of a subject protein in real time.

主权项

1. A system for measuring or characterizing a protein in a sample in real time during electrophoresis, the system comprising:
a dual-labeled protein standard comprising:
a plurality of purified proteins of differing molecular weight;wherein at least a portion of the purified proteins are covalently labeled with a first fluorescent dye, said first fluorescent dye having a first excitation energy and a first emission spectrum; andwherein at least two of the purified proteins are covalently labeled with a second fluorescent dye, said second fluorescent dye having a second excitation energy and a second emission spectrum; a sample comprising a protein to be characterized, said protein being covalently labeled with said second fluorescent dye; and a means of simultaneously detecting said first and second fluorescent dye; wherein the first fluorescent dye and the second fluorescent dye are different; wherein the first excitation energy and the second excitation energy are substantially similar; wherein the purified proteins labeled with the first fluorescent dye comprise a molecular weight standard, and wherein the purified proteins labeled with the second fluorescent dye comprise a mass standard.