Abstract: Oxidative stress (OS) is associated with a wide variety of diseases and disorders. Detection of oxidative stress in living systems typically relies on fluorescent probes for reactive oxygen species (ROS), which is challenging because of their short life span and high reactivity. OS-induced biomolecule carbonylation is a stable modification that also possesses a chemically reactive functional group, and may be detected with a hydrazine, alkoxyamine or hydrazide-containing probe, in a hydrazone or oxime-forming reaction, that does not require strong acid catalysis or nucleophilic catalysis with an aromatic amine. Fluorophores possessing hydrazine, alkoxyamine or hydrazide functional groups can undergo reaction with carbonylated biomolecules in live cells, fixed cells, and tissue sample, and these products can be observed using fluorescence microscopy.

Abstract: A reaction method comprising combining a carbonyl-substituted arylboronic acid or ester and an ?-effect amine in aqueous solution at a temperature between about ?5 C to 55 C, and a pH between 2 and 8 to produce an adduct. A process is also provided comprising: contacting a boron compound having a boron atom bonded to a sp2 hybridized carbon conjugated with a cis-carbonyl, the boron having at least one labile substituent, with an ?-effect amine, in a solvent for a time sufficient to form an adduct, which may proceed to further products.

Abstract: A technique is provided to visualize microtubules in live cells that does not require genetic manipulation or microinjection. Moreover, this method also avoids perturbation of the endogenous microtubule network that occurs with taxol treatment. This technique exploits tyrosination and detyrosination of tubulin, a posttranslational modification cycle specific to the C-terminus of ?-tubulin. Specifically, cells are grown in medium supplemented with a tyrosine derivative possessing a reactive functional group. The cellular enzyme tubulin tyrosine ligase attaches the unnatural amino acid to a single site on tubulin. Addition of fresh medium containing a suitably derivatized fluorophore then yields fluorescent tubulin, which incorporate into cellular microtubules. Importantly, the tubulin labeling approach demonstrated here does not detrimentally affect microtubule network or cell morphology. Thus we present a simple, robust labeling technique that allows microscopic analysis of microtubules in live cells.

Abstract: A reaction method comprising combining a carbonyl-substituted arylboronic acid or ester and an ?-effect amine in aqueous solution at a temperature between about ?5 C to 55 C, and a pH between 2 and 8 to produce an adduct. A process is also provided comprising: contacting a boron compound having a boron atom bonded to a sp2 hybridized carbon conjugated with a cis-carbonyl, the boron having at least one labile substituent, with an ?-effect amine, in a solvent for a time sufficient to form an adduct, which may proceed to further products.

Abstract: A reaction method comprising combining a carbonyl-substituted arylboronic acid or ester and an ?-effect amine in aqueous solution at a temperature between about ?5 C to 55 C, and a pH between 2 and 8 to produce an adduct. A process is also provided comprising: contacting a composition having a boron atom bonded to a sp2 hybridized carbon, the boron having at least one labile substituent, conjugated with a cis-carbonyl, with an ?-effect amine, in an aqueous medium for a time sufficient to form an adduct, which may proceed to further products.

Abstract: A reaction method comprising combining a carbonyl-substituted arylboronic acid or ester and an ?-effect amine in aqueous solution at a temperature between about ?5 C to 55 C, and a pH between 2 and 8 to produce an adduct. A process is also provided comprising: contacting a composition having a boron atom bonded to a sp2 hybridized carbon, the boron having at least one labile substituent, conjugated with a cis-carbonyl, with an ?-effect amine, in an aqueous medium for a time sufficient to form an adduct, which may proceed to further products.

Abstract: A technique is provided to visualize microtubules in live cells that does not require genetic manipulation or microinjection. Moreover, this method also avoids perturbation of the endogenous microtubule network that occurs with taxol treatment. This technique exploits tyrosination and detyrosination of tubulin, a posttranslational modification cycle specific to the C-terminus of ?-tubulin. Specifically, cells are grown in medium supplemented with a tyrosine derivative possessing a reactive functional group. The cellular enzyme tubulin tyrosine ligase attaches the unnatural amino acid to a single site on tubulin. Addition of fresh medium containing a suitably derivatized fluorophore then yields fluorescent tubulin, which incorporate into cellular microtubules. Importantly, the tubulin labeling approach demonstrated here does not detrimentally affect microtubule network or cell morphology. Thus we present a simple, robust labeling technique that allows microscopic analysis of microtubules in live cells.