System for rapid electrophoresis binding method and related kits and compositions

Abstract

An improved staining method and system is described for staining a biopolymer such as a peptide, a protein, an RNA, a DNA, an oligosaccharide or a complex containing a peptide, a protein, an RNA, a DNA, or an oligosaccharide in a matrix. The method includes the step of moving a staining reagent into the matrix using an electric force. The staining time can be dramatically reduced relative to conventional technologies. The improved staining method can particularly be used, for example, to stain proteins after gel separation. Other related methods, related kits and related systems for carrying out the staining method are also described.

We claim: 
     
         1 . A system for staining a target molecule with a staining reagent, the system comprising:
 (i) a matrix to be embedded with a target molecule to form a matrix embedded with the target molecule;   (ii) a first solid porous material for forming a first solid porous material absorbed with a staining solution comprising the staining reagent, wherein the first solid porous material absorbed with a staining solution is to be placed in contact with the matrix embedded with the target molecule;   (iii) a pair of electrodes for applying an electric force sufficient to move the staining reagent from the first solid porous material into the matrix embedded with the target molecule, thereby staining the target molecule, and to move excess free staining reagent out of the matrix, thereby destaining the matrix; and   (iv) optionally, one or more additional solid porous materials absorbed with one or more buffer solutions.   
     
     
         2 . The system according to  claim 1 , wherein the matrix embedded with the target molecule and the first solid porous material are interposed between the pair of electrodes, such that the electric force moves the staining reagent in a direction such that the excess free staining reagent enters the matrix and travels the least distance in the matrix before leaving the matrix as compared to the other directions. 
     
     
         3 . The system according to  claim 1 , wherein the matrix is selected from the group consisting of an agarose gel, a polyacrylamide gel, and any porous material capable of being embedded with a target molecule; the target molecule is a biopolymer selected from the group consisting of a peptide, a protein, an RNA, a DNA, an oligosaccharide, and a complex thereof; and the staining reagent is a charged molecule selected from the group consisting of an organic or inorganic reagent, a dye or a dye-labeled reagent, a fluorescent molecule, a metal-complex dye, a radioactive or radioactive-labeled reagent, an antibody or an antibody-based reagent, a labeled peptide or ligand, and a labeled polynucleotide. 
     
     
         4 . The system according to  claim 1 , wherein the first porous material is further absorbed with an electrolytic buffer. 
     
     
         5 . The system according to  claim 1  being an automated system further comprising an automated control unit connected to the pair of electrodes, the automated control unit comprising one or more components selected from the group consisting of a display panel, a power supply, and a power switch. 
     
     
         6 . The system according to  claim 5 , wherein the automated control unit is connected to a computer interface that directs the automated control unit to control one or more parameters selected from the group consisting of run time, voltage, current, number of run cycles, analysis and processing of input data, and run start, run stop, and run pause commands. 
     
     
         7 . A system for staining a target molecule with a staining reagent, wherein the system comprises, in the following order:
 (i) a first solid porous material absorbed with a staining solution comprising the staining reagent;   (ii) a matrix embedded with the target molecule;   (iii) a second solid porous material absorbed with an electrolytic buffer; and   (iv) a pair of electrodes for applying an electric force sufficient to move the staining reagent from the first solid porous material into the matrix embedded with the target molecule, thereby staining the target molecule, and to move excess free staining reagent out of the matrix, thereby destaining the matrix;   wherein the pair of electrodes are to be connected to the first solid porous material and second solid porous material, respectively.   
     
     
         8 . The system according to  claim 7 , wherein the matrix embedded with the target molecule and the first solid porous material are interposed between the pair of electrodes, such that the electric force moves the staining reagent in a direction such that the excess free staining reagent enters the matrix and travels the least distance in the matrix before leaving the matrix as compared to the other directions. 
     
     
         9 . The system according to  claim 7 , wherein the matrix is selected from the group consisting of an agarose gel, a polyacrylamide gel, and any porous material capable of being embedded with a target molecule; the target molecule is a biopolymer selected from the group consisting of a peptide, a protein, an RNA, a DNA, an oligosaccharide, and a complex thereof; and the staining reagent is a charged molecule selected from the group consisting of an organic or inorganic reagent, a dye or a dye-labeled reagent, a fluorescent molecule, a metal-complex dye, a radioactive or radioactive-labeled reagent, an antibody or an antibody-based reagent, a labeled peptide or ligand, and a labeled polynucleotide. 
     
     
         10 . The system according to  claim 7 , wherein the matrix is a polyacrylamide gel embedded with a protein or peptide. 
     
     
         11 . The system of  claim 7 , further comprising a third solid porous material absorbed with a washing buffer, wherein the third solid porous material is placed adjacent to the first solid porous material, and the pair of electrodes are connected to the third solid porous material absorbed with the washing buffer and the second solid porous material absorbed with the electrolytic buffer. 
     
     
         12 . The system according to  claim 11 , wherein the staining solution comprises, per liter of the staining solution, about 10 grams to about 300 grams acetic acid, about 10 grams to about 500 grams isopropanol, about 0.1 grams to about 10 grams EDTA, about 0.1 gram to about 10 grams Tris, about 0.1 gram to 5 grams Coomassie Blue, about 0.1 gram to about 10 grams Na 2 HPO 4 , and about 0.1 gram to about 10 grams NaCl, and the pH being in the range of about 2.0 to about 11.0; the electrolytic buffer comprises, per liter of electrolytic buffer, about 1 gram to about 50 grams sodium phosphate, monobasic, and the pH being in the range of about 2.0 to about 10.0; and the washing buffer comprises, per liter of washing buffer, about 10 grams to about 300 grams acetic acid, about 10 grams to about 500 grams isopropanol, about 0.1 grams to about 10 grams EDTA, about 0.1 gram to about 10 grams Tris, about 0.1 gram to about 10 grams Na 2 HPO 4 , and about 0.1 gram to about 10 grams NaCl, and the pH being in the range of about 2.0 to about 11.0. 
     
     
         13 . The system according to  claim 7  being an automated system further comprising an automated control unit connected to the pair of electrodes, the automated control unit comprising one or more components selected from the group consisting of a display panel, a power supply, and a power switch. 
     
     
         14 . The system according to  claim 13 , wherein the automated control unit is connected to a computer interface that directs the automated control unit to control one or more parameters selected from the group consisting of run time, voltage, current, number of run cycles, analysis and processing of input data, and run start, run stop, and run pause commands. 
     
     
         15 . The system of  claim 1 , further comprising a kit for staining the target molecule embedded in the matrix with the staining reagent, the kit comprising:
 (i) the staining solution comprising the staining reagent;   (ii) a plurality of solid porous materials;   (iii) optionally one or more buffer solutions;   (iv) optionally one or more matrices for embedding a target molecule; and   (v) instructions for using the staining solution and optionally the buffer solutions for staining the biopolymer in the matrix utilizing an electric force applied to the staining reagent,   
       wherein the staining solution and buffer solutions are optionally pre-absorbed in separate solid porous materials. 
     
     
         16 . A system for staining a target molecule with a staining reagent, the system comprising:
 (i) a matrix to be embedded with a target molecule to form a matrix embedded with the target molecule;   (ii) a staining reservoir for receiving the matrix embedded with the target molecule;   (iii) one or more pumps to be connected to the staining reservoir for pumping a staining solution comprising a staining reagent into and out of the staining reservoir, and for pumping a washing buffer into and out of the staining reservoir; and   (iv) a pair of electrodes for applying an electric force sufficient to move the staining reagent from the staining solution that is pumped into the staining reservoir into the matrix embedded with the target molecule, thereby staining the target molecule, and to move excess free staining reagent out of the matrix into the washing buffer that is pumped into the staining reservoir, thereby destaining the matrix.   
     
     
         17 . The system according to  claim 16 , further comprising a waste collection unit to be connected to the staining reservoir for receiving the staining solution and destaining buffer that is pumped out of the staining reservoir. 
     
     
         18 . The system according to  claim 16 , wherein the matrix is selected from the group consisting of an agarose gel, a polyacrylamide gel, and any porous material capable of being embedded with a target molecule; the target molecule is a biopolymer selected from the group consisting of a peptide, a protein, an RNA, a DNA, an oligosaccharide, and a complex thereof; and the staining reagent is a charged molecule selected from the group consisting of an organic or inorganic reagent, a dye or a dye-labeled reagent, a fluorescent molecule, a metal-complex dye, a radioactive or radioactive-labeled reagent, an antibody or an antibody-based reagent, a labeled peptide or ligand, and a labeled polynucleotide. 
     
     
         19 . The system according to  claim 16  being an automated system further comprising an automated control unit connected to the pair of electrodes, the automated control unit comprising one or more components selected from the group consisting of a display panel, a power supply, and a power switch, wherein the automated control unit is further connected to a computer interface that directs the automated control unit to control one or more parameters selected from the group consisting of run time, voltage, current, number of run cycles, analysis and processing of input data, run start, run stop, and run pause commands, and pump flow rate. 
     
     
         20 . The system according to  claim 16 , wherein the staining solution comprises, per liter of the staining solution, about 10 grams to about 300 grams acetic acid, about 10 grams to about 500 grams isopropanol, about 0.1 grams to about 10 grams EDTA, about 0.1 gram to about 10 grams Tris, about 0.1 gram to 5 grams Coomassie Blue, about 0.1 gram to about 10 grams Na 2 HPO 4 , and about 0.1 gram to about 10 grams NaCl, and the pH being in the range of about 2.0 to about 11.0; and the washing buffer comprises, per liter of washing buffer, about 10 grams to about 300 grams acetic acid, about 10 grams to about 500 grams isopropanol, about 0.1 grams to about 10 grams EDTA, about 0.1 gram to about 10 grams Tris, about 0.1 gram to about 10 grams Na 2 HPO 4 , and about 0.1 gram to about 10 grams NaCl, and the pH being in the range of about 2.0 to about 11.0.