Abstract: The invention provides barcode libraries and methods of making and using them including obtaining a plurality of nucleic acid constructs in which each construct comprises a unique N-mer and a functional N-mer and segregating the constructs into a fluid compartments such that each compartment contains one or more copies of a unique construct. The invention further provides methods for digital PCR and for use of barcode libraries in digital PCR.

Abstract: A method of creating a protective coating on an alkali metal hydroxide-containing solid is provided. The method includes providing carbon dioxide to an alkali metal hydroxide-containing solid and allowing the alkali metal hydroxide and carbon dioxide to react thereby forming a carbonate or bicarbonate-containing layer on the exterior of the solid wherein the carbonate or bicarbonate-containing layer is non-hygroscopic and water soluble, and wherein greater than 80% of the hydroxide in the hydroxide-containing solid does not react with the carbon dioxide, and further wherein the alkali metal hydroxide-containing solid is substantially free of lithium hydroxide. A method of testing for the presence of carbonate-containing coating on an alkali metal hydroxide containing solid is also provided. The method includes exposing the coated solid to 95 weight percent ethanol, collecting the ethanol effluent and testing the effluent for alkali metal hydroxide.

Abstract: Methods and genetic constructs are provided for detecting the binding of nuclear hormone receptors to a coactivator/corepressor. The methods employ enzyme fragment complementation using fragments of ?-galactosidase as the detection system. Cells are transformed to express the large fragment of ?-galactosidase fused to a member of the complex with NHR for initiation of transcription and have it localized in the nucleus and to express the small fragment of ?-galactosidase fused to the nuclear hormone receptor for binding to the member upon stimulation with a ligand.

Abstract: A method of detecting first and second analytes includes providing a mixture containing first analytes and second analytes; adding microparticles of a first size coated with first competitive inhibitors that compete with the first analytes for binding to first antibodies to the first analytes, and adding microparticles of a second size coated with second competitive inhibitors that compete with the second analytes for binding to first antibodies to the second analytes, adding second antibodies specific to the first antibodies to the first analytes and second antibodies specific to the first antibodies to the second analytes, wherein the second antibodies specific to the first antibodies to the first analytes are labeled with a first fluorescent moiety, and the second antibodies specific to the first antibodies to the second analytes are labeled with a second fluorescent moiety

Abstract: Methods for detecting phosphorylation of receptor tyrosine kinases (“RTKs”) upon activation and the modulation of activation by a candidate compound are provided. The method employs cells comprising two fusion products: (1) an RTK fused to a small fragment of ?-galactosidase and (2) a phosphotyrosine binding peptide fused to the large fragment of ?-galactosidase, where the 2 fragments weakly complex to form an active enzyme, and optionally a construct for a cytosolic RTK phosphorylating kinase, when the RTK does not autophosphoryate. To detect phosphorylation a ?-galactosidase substrate is added to the cells, whereby product formation indicates the occurrence of phosphorylation.

Abstract: Truncated fragments of the small fragment of ?-galactosidase are provided that have low affinity for the large fragment of ?-galactosidase and provide for robust signals when two fusion proteins are complexed due to the binding of the proteins to which the ?-galactosidase fragments are fused. The truncated fragments do not interfere with the complexing of the two proteins and allow for the two proteins to function and be responsive to candidate compounds that affect complex formation.

Abstract: Methods for detecting phosphorylation of receptor tyrosine kinases (“RTKs”) upon activation are provided. The method employs cells comprising two fusion products: (1) an RTK fused to a small fragment of ?-galactosidase and (2) a phosphotyrosine binding peptide fused to the large fragment of ?-galactosidase, where the 2 fragments weakly complex to form an active enzyme, and optionally a construct for a cytosolic RTK phosphorylating kinase, when the RTK does not autophosphoryate. To detect phosphorylation a ?-galactosidase substrate is added to the cells, whereby product formation indicates the occurrence of phosphorylation.

Abstract: A method of detecting analytes includes providing a mixture containing analytes to be analyzed and microparticles coated with competitive inhibitors of the analytes. First antibodies are reacted with the analytes and the competitive inhibitors under competitive binding conditions. The first antibodies bound to the competitive inhibitors are measured using flow cytometry.

Abstract: Methods and genetic constructs are provided for detecting the binding of nuclear hormone receptors to a coactivator/corepressor. The methods employ enzyme fragment complementation using fragments of ?-galactosidase as the detection system. Cells are transformed to express the large fragment of ?-galactosidase fused to a member of the complex with NHR for initiation of transcription and have it localized in the nucleus and to express the small fragment of ?-galactosidase fused to the nuclear hormone receptor for binding to the member upon stimulation with a ligand.

Abstract: Sensitive assays for candidate compounds affecting GPCR activity are provided using a cell containing fusion proteins comprising a first fusion protein comprising (a) a target GPCR fused to a small fragment of ?-galactosidase through a linker comprising a phosphorylation site or (b) a GPCR or a protein of interest, where the GPCR and protein of interest form a complex and one of them is fused to the small fragment of ?-galactosidase; and a second fusion protein comprising arrestin fused to a large fragment of ?-galactosidase. In (a), the affinity of the small and large fragments is optimized based on the background to signal ratio and the absolute signal observed. The assay is performed using a ?-galactosidase substrate that provides a detectable optical signal.

Abstract: Methods for detecting phosphorylation of receptor tyrosine kinases (“RTKs”) upon activation are provided. The method employs cells comprising two fusion products: (1) an RTK fused to a small fragment of ?-galactosidase and (2) a phosphotyrosine binding peptide fused to the large fragment of ?-galactosidase, where the 2 fragments weakly complex to form an active enzyme, and optionally a construct for a cytosolic RTK phosphorylating kinase, when the RTK does not autophosphoryate. To detect phosphorylation a ?-galactosidase substrate is added to the cells, whereby product formation indicates the occurrence of phosphorylation.

Abstract: Truncated fragments of the small fragment of ?-galactosidase are provided that have low affinity for the large fragment of ?-galactosidase and provide for robust signals when two fusion proteins are complexed due to the binding of the proteins to which the ?-galactosidase fragments are fused. The truncated fragments do not interfere with the complexing of the two proteins and allow for the two proteins to function and be responsive to candidate compounds that affect complex formation.

Abstract: A piston apparatus for use in a controlled deflection roll includes a rectilinear-shaped piston having at least one side seal extending longitudinally in a slot in each side wall and an end seal in each end of the piston extending between the side seals. The end seals have at least one pair of intersecting surfaces at each end thereof which cooperate with a corresponding pair of surfaces of each side seal to move longitudinally relative to the piston and side seals to maintain a continuous seal peripherally about the piston. The continuous seal engages the side walls of a channel within the support shaft of the controlled deflection roll and a non-movable seal surface in each of two, opposed dams at either end of the channel in the support shaft. The side and end seals move with the piston relative to the support shaft and seal surfaces on the end dams.