Simultaneous identification of 36 mutations in KRAS codons 61and 146, BRAF, NRAS, and PIK3CAin a single reaction by multiplex assay kit

δ-Catenin is overexpressed in human cancers, including prostate, breast, lung, and ovarian cancers. Therefore, detection of δ-catenin level in patient specimens can be used as a diagnostic marker for the cancer screening. In laboratories, δ-catenin levels have been analyzed by western blot, which requires multiple procedures and is incapable of multiplex analysis of a target protein from a single reaction. In this study, we aimed to develop δ-catenin antibody-conjugated magnetic beads that can be used for quantitation of δ-catenin by bead-based multiplex assay. δ-catenin level from HEK293T and CWR22Rv-1 cell lysates was quantified to determine whether the antibody-conjugatedmagnetic bead can be used for the detection of trace amounts of δ-catenin. Our results indicate that analysis with 1 μg of CWR22Rv-1 and HEK293T cell lysates showed 918.67±103.8 and 874.33±37.21 MFI values, respectively. In conclusion, the results suggest that the procedure for detection of δ-catenin level through the antibody-conjugated magnetic beads provides a sensitive and costeffective solution for screening from patient specimens.

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A high multiplex assay to quantify copy number variation in the FGFR1, 2, 3 genes and chromosome 11q in a single reaction

European Journal of Cancer ◽

10.1016/s0959-8049(16)32998-7 ◽

2016 ◽

Vol 69 ◽

pp. S134

Author(s):

K. Madanahally Divakar ◽

M. Boehme ◽

K. Korn

Keyword(s):

Copy Number Variation ◽

Copy Number ◽

Multiplex Assay ◽

Number Variation ◽

Single Reaction

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Analyzing reactions of single mechanoenzyme molecules by light microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100122538 ◽

1992 ◽

Vol 50 (1) ◽

pp. 426-427

Author(s):

Jeff Gelles

Keyword(s):

Image Processing ◽

Reaction Mechanisms ◽

Transient State ◽

Nanometer Scale ◽

Reaction Intermediates ◽

Enzyme Molecule ◽

Directed Movement ◽

Enzyme Molecules ◽

Individual Enzyme ◽

Single Reaction

Mechanoenzymes are enzymes which use a chemical reaction to power directed movement along biological polymer. Such enzymes include the cytoskeletal motors (e.g., myosins, dyneins, and kinesins) as well as nucleic acid polymerases and helicases. A single catalytic turnover of a mechanoenzyme moves the enzyme molecule along the polymer a distance on the order of 10−9 m We have developed light microscope and digital image processing methods to detect and measure nanometer-scale motions driven by single mechanoenzyme molecules. These techniques enable one to monitor the occurrence of single reaction steps and to measure the lifetimes of reaction intermediates in individual enzyme molecules. This information can be used to elucidate reaction mechanisms and determine microscopic rate constants. Such an approach circumvents difficulties encountered in the use of traditional transient-state kinetics techniques to examine mechanoenzyme reaction mechanisms.

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