Submillimeter-scale surface gradients of immobilized protein ligands
Date
2009
Authors
Walker, R.A.
Cunliffe, V.T.
Whittle, J.D.
Steele, D.A.
Short, R.D.
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Journal article
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Langmuir, 2009; 25(8):4243-4246
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Abstract
We describe a method to produce antibody-captured ligand gradients over biologically relevant distances (hundreds of micrometers) whereby the ligand density and gradient shape may be tailored. Separation of the ligand from the solid-phase surface ensures that the biological activity of the ligand remains unaffected by immobilization. Our method involves the use of a plasma-masking method to generate a surface chemical gradient on a glass substrate to which the 9E10 antibody is covalently coupled. This antibody captures myc-tagged biomolecules. In our example, the antibody is then used to immobilize a gradient of the intercellular signaling molecule delta-like-1 (Dll1). To visualize the gradient of Dll1, we have used the multistep approach of binding with rabbit anti-Dll1 primary antibody and then adding colloidal-gold-conjugated secondary antibody.
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Copyright 2009 American Chemical Society