Compositions and Methods for Improving Resolution of Biomolecules Separated on Polyacrylamide Gels

Abstract

Gels, such as polyacrylamide gels, are provided that include linear polyacrylamide in the stacking gel. Native gels that include linear polyacrylamide in the stacker can be used to separate biomolecular complexes, such as protein complexes. Gel cassettes in which the gap width between front and back plates does not vary by more than 5% at the upper edge of the cassette are also provided. The gel cassettes can be used for electrophoretic separation of proteins and protein complexes on native gels, such as native gels that include linear polyacrylamide in the stacker. The native gels can have multiple wells for electrophoresing at least one sample and/or at least one molecular weight standard.

1 . A polyacrylamide gel for the separation of biomolecules, comprising a stacking gel and a separating gel, wherein the stacking gel comprises linear polyacrylamide.  
     
     
         2 . The polyacrylamide gel of  claim 1 , wherein the acrylamide concentration of the stacking gel is between 2% and 6%.  
     
     
         3 . The polyacrylamide gel of  claim 2 , wherein the acrylamide concentration of the stacking gel is between 2.5% and 5%.  
     
     
         4 . The polyacrylamide gel of  claim 2 , wherein the linear acrylamide concentration of the stacking gel is from 0.005% to 1%.  
     
     
         5 . The polyacrylamide gel of  claim 4 , wherein the linear acrylamide concentration of the stacking gel is from 0.01% to 0.5%.  
     
     
         6 . The polyacrylamide gel of  claim 5 , wherein the linear acrylamide concentration of the stacking gel is from 0.02% to 0.1%.  
     
     
         7 . The polyacrylamide gel of  claim 1 , wherein the separating gel does not comprise linear acrylamide.  
     
     
         8 . The polyacrylamide gel of  claim 1 , wherein the polyacrylamide gel is a denaturing gel.  
     
     
         9 . The polyacrylamide gel of  claim 8 , wherein the polyacrylamide gel comprises SDS.  
     
     
         10 . The polyacrylamide gel of  claim 1 , wherein the polyacrylamide gel is a nondenaturing gel.  
     
     
         11 . The polyacrylamide gel of  claim 10 , wherein the nondenaturing gel is a gradient gel.  
     
     
         12 . The polyacrylamide gel of  claim 10 , wherein the non-denaturing gel is a Blue Native Gel.  
     
     
         13 . A method of separating biomolecules on an electrophoresis gel, comprising: 
 applying one or more samples comprising one or more biomolecules to an acrylamide gel comprising a stacking gel portion and a separating gel portion, wherein the stacking gel portion comprises linear polyacrylamide; and    electrophoretically separating one or more biomolecules on the gel.    
     
     
         14 . The method of  claim 13 , wherein the separating gel portion does not comprise linear acrylamide.  
     
     
         15 . The method of  claim 13 , wherein the electrophoresis gel is a denaturing gel.  
     
     
         16 . The method of  claim 13 , wherein the electrophoresis gel is a nondenaturing gel.  
     
     
         17 . The method of  claim 16 , wherein the nondenaturing gel is a Blue Native gel.  
     
     
         18 . The method of  claim 16 , wherein the nondenaturing gel is a gradient gel.  
     
     
         19 . The method of  claim 13 , further comprising applying one or more molecular weight marker sets to the electrophoresis gel.  
     
     
         20 . The method of  claim 19 , further comprising estimating or calculating the molecular weight of one or more biomolecules or biomolecular complexes on the electrophoresis gel.  
     
     
         21 - 41 . (canceled)