Bio-rad Laboratories, Inc.

Other

Thomas R Berkelman

Lee Olech

Cory Panattoni

Xuemei Yang

Craig Rowell

Shane Petersen

John Walker, Ii

Sean Cater

Lisandra Martin

Rhiannon Jones

Christopher Belisle

William Strong

Clayton T McKee

Praveena D Garimella

John Walker Ii

Clayton T Mckee

Garimella Praveena

Polyacrylamide gels that offer high resolution in protein separations and are more stable relative to hydrolysis than conventional polyacrylamide gels that rely on Tris or Tris-Bis as buffering agents are made by incorporating triethanolamine in place of most or all of the Tris or Tris-Bis.

Hydrolysis-resistant polyacrylamide gels

Conjugates of 1,3-bis(1,4,7-triazacyclonon-1-yl)-2-hydroxypropanes with a variety of conjugating members are used in the formation of dinuclear metal complexes which bind to phosphate esters. By virtue of their conjugated forms, the complexes are incorporated into chromatographic media, affinity binding reagents, and dyes, which make the complexes useful in a wide range of assays, separations, and purifications. In addition, dinuclear metal complexes of 1,3-bis(1,4,7-triazacyclonon-1-yl)-2-hydroxypropanes that are not so conjugated are used in the detection of phosphate esters of biological species by either MALDI-TOF mass spectrometry or by dye displacement.

Conjugates of 1,4,7-triazacyclononanes, dinuclear metal complexes of such conjugates, and methods of use for both 1,4,7-triazacyclononanes and conjugate

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.

Protein detection using modified cyclodextrins

Conjugates of 1,4,7-triazacyclononanes, dinuclear metal complexes of such conjugates, and methods of use for both 1,4,7-triazacyclononanes and conjugates

Colloidal formulation for staining proteins and methods of their use are provided.

Colloidal Coomassie stain

Protein dyes whose molecular structure is that of a coumarin moiety coupled to a quaternary ammonium heterocycle through a vinyl or polyvinyl linkage demonstrate the ability to associate with proteins in a non-covalent, non-specific manner at low pH, where the associated form displays a significantly higher fluorescence emission than the unassociated form. This makes the dyes useful as selective labels for proteins at the low pH and eliminates the need for the removal of extraneous components from the medium in which the proteins reside prior to detection.

Coumarin-based cyanine dyes for non-specific protein binding

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Oakland, CA, United States of America

Thomas R Berkelman