公开(公告)号：EP2986646A4

公开(公告)日：2017-04-12

申请号：EP14834424

申请日：2014-08-08

Applicant: BIO-RAD LABORATORIES INC

Inventor： MCKEE CLAYTON T ,  STRONG WILLIAM ,  OLECH LEE ,  BERKELMAN THOMAS ,  BELISLE CHRISTOPHER

IPC: G01N33/58 ,  G01N27/447 ,  G01N33/68

CPC classification number: G01N33/5306 ,  G01N27/44747 ,  G01N33/582 ,  G01N33/6803 ,  G01N2400/18

Abstract: A method is provided for detecting a protein using a cyclodextrin covalently linked to at least one label. The cyclodextrin can associate with the protein by sequestering an aromatic amino acid side-chain of the protein in its hydrophobic cavity. After contacting the protein with the cyclodextrin, the label can be detected directly or can undergo a chemical interaction with a reagent to form a detectable product. The label can include an indole moiety, which can react with a halo-substituted organic compound upon exposure to UV light and thereby be rendered fluorescent. Alternatively, the label can include a biotin moiety, which can bind to a binding partner such as avidin, or variants thereof, to form a detectable molecular complex. A labeled cyclodextrin can be used in the present methods to detect a protein of interest in an electrophoresis gel or on a blotting membrane. Aromatic amino acid residues of the protein, in particular tryptophan, remain protected from chemical modification due to sequestration by the cyclodextrin, making these methods compatible with downstream applications that require intact protein. Also provided herein are compositions, kits, and electrophoresis gels for use in detecting proteins.