Hemicyanine dyes
AssigneeLI-COR, Inc.
InventorsSilvano Sanchini, Rose Skopp, Vassil Elitzin, Nisha Padhye, Teresa Urlacher
Life of this patent
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Abstract
The present disclosure embodies compounds, methods, and kits that are useful in connection with the detection of analytes of all types (e.g., biological molecules such as proteins, organic molecules, natural or synthetic molecules). The disclosure is particularly applicable to detection of proteins and nucleic acids using all types of membrane-based assays by techniques such as Western blotting, Dot blotting, Southern blotting, and Northern blotting.
Claims50
A compound having Formula I:
- wherein R1, R2, R3, R4, R5 and R6 are each a member independently selected from the group consisting of hydrogen, alkyl, alkenyl, halo, hydroxyl, alkoxy, amino, cyano, carboxyl, carboxylate, alkoxycarbonyl, amido, sulfonato, alkoxyalkyl, carboxyalkyl, alkoxycarbonylalkyl, sulfonatoalkyl, a functional group for conjugation to a biomolecule, and R7B, wherein R7 is the resultant attachment between the compound and a biomolecule;
- B is a biomolecule;
- x, y and z are each an integer independently selected from 1-10;
- and
- X is hydrogen or a metal cation, wherein said compound has a balanced charge, wherein in Formula I, at least one of R1, R2, R3, R4, R5 and R6 is a functional group for conjugation or is R7B.
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A compound having Formula I(a):
- wherein R1, R2, R3, R4, R5 and R6 are each a member independently selected from the group consisting of hydrogen, alkyl, alkenyl, halo, hydroxyl, alkoxy, amino, cyano, carboxyl, carboxylate, alkoxycarbonyl, amido, sulfonato, alkoxyalkyl, carboxyalkyl, alkoxycarbonylalkyl, and sulfonatoalkyl;
- x, y and z are each an integer independently selected from 1-10;
- and
- X is hydrogen or a metal cation, wherein said compound has a balanced charge.
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A method for detecting a target biomolecule in a sample, said method comprising:
- providing a sample that is suspected of containing a target analyte;
- providing a compound of Formula I or Formula I(a) conjugated to a capture molecule, wherein the capture molecule is capable of interacting with the target analyte;
- contacting the sample with the capture molecule and the compound of Formula I or Formula I(a) under conditions in which the capture molecule can bind to the target analyte if present;
- applying a light source to the sample that excites the compound of Formula I or Formula I(a);
- and detecting whether light is emitted from the conjugated compound of Formula I or Formula I(a).
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A method for detecting and or staining a protein, the method comprising:
- contacting a staining mixture that contains a compound or dye of Formula I or Formula I(a) with a protein to form a combined mixture;
- incubating the combined mixture for a time sufficient for the compound or dye of Formula I to associate with the protein to form a dye complex that gives an optical response upon illumination;
- illuminating the dye-protein complex;
- and
- detecting the optical response.
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A kit for staining a protein in a sample, the kit comprising:
- a) a compound or dye or Formula I or Formula I(a);
- and
- b) instructions for combining said compound of Formula I or Formula I(a) with a sample containing or thought to contain protein;
- said instructions comprising:
- i) combining a sample that is thought to contain a protein with a staining mixture that contains said compound of Formula I or Formula I(a) to form a combined mixture;
- and
- ii) incubating the combined mixture for a time sufficient for the dye in the staining mixture to associate with the protein in the sample mixture to form a dye-protein complex that gives a detectable optical response upon illumination.
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Dependencies are inferred from the claim text.