Targeted delivery of reagents to spots on a planar support through patterned transfer sheets

Abstract

Molecular species that are immobilized in discrete locations on a planar support such as protein bands on a gel or a blotting membrane or species applied in dots or spots on a membrane are reacted with binding reagents that are applied through a porous hydrophilic transfer sheet placed over the planar support, the sheet having at least one region that is laterally bordered by a barrier with the binding reagent retained within the bordered region. The bordered region is placed directly over an area on the planar support where the molecular species are expected to reside if they are present on the support. The binding reagent is then delivered into the support to contact the species. Targeted delivery of the binding reagent is thus achieved with improved efficiency.

1 - 10 . (canceled) 
     
     
         11 . A method of making an aqueous-based binding reagent transfer sheet, wherein the aqueous-based binding reagent is reversibly immobilized in a laterally delimited region that is bordered by a barrier, the method comprising
 providing a porous hydrophilic transfer sheet;   contacting the transfer sheet with a silanization reagent to form a laterally delimited region bordered by a barrier of silanized transfer sheet; and   contacting the laterally delimited region with an aqueous solution containing a binding reagent,   thereby making transfer sheet with an aqueous-based binding reagent that is reversibly immobilized in a laterally delimited region that is bordered by a barrier.   
     
     
         12 . The method of  claim 11  wherein the silanization reagent is trimethylchlorosilane. 
     
     
         13 . The method of  claim 11  wherein the binding reagent is an antibody. 
     
     
         14 . The method of  claim 11  wherein the method further comprises contacting the laterally delimited region with a second aqueous solution containing a second binding reagent. 
     
     
         15 . The method of  claim 14  wherein the second binding reagent is an antibody. 
     
     
         16 . The method of  claim 11  wherein the porous hydrophilic transfer sheet comprises cellulose. 
     
     
         17 . The method of  claim 11  wherein the transfer sheet is masked before contacting with the silanization reagent. 
     
     
         18 . A kit comprising a container and 1, 2, 3, 4, or more porous hydrophilic sheets residing therein, the porous hydrophilic sheets each having 1, 2, 3, 4, or more laterally delimited regions therein that are bordered by a hydrophobic or impermeable barrier. 
     
     
         19 . The kit of  claim 18 , wherein the porous hydrophilic sheets each have 1, 2, 3, 4, or more binding reagents reversibly immobilized therein. 
     
     
         20 . The kit of  claim 18 , wherein the laterally delimited regions of the porous hydrophilic sheets are substantially the width of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more lanes in an electroblot. 
     
     
         21 . The kit of  claim 18 , wherein the container is configured to protect the porous hydrophilic sheets from light and/or moisture. 
     
     
         22 . The kit of  claim 18 , further comprising a transfer buffer.