The Experts below are selected from a list of 177 Experts worldwide ranked by ideXlab platform

Tomoji Kawai - One of the best experts on this subject based on the ideXlab platform.

Xi Chen - One of the best experts on this subject based on the ideXlab platform.

Hiroyuki Tanaka - One of the best experts on this subject based on the ideXlab platform.

Lee Josephson - One of the best experts on this subject based on the ideXlab platform.

  • Fluorescein Isothiocyanate–hapten immunoassay for determination of peptide–cell interactions
    Analytical Biochemistry, 2004
    Co-Authors: Kimberly A. Kelly, Fred Reynolds, Ralph Weissleder, Lee Josephson
    Abstract:

    Abstract We have developed a Fluorescein Isothiocyanate (FITC)–hapten immunoassay, where a FITC-labeled peptide binding to a cell is assayed as the amount of immunoreactive Fluorescein present in a cell lysate. An antiFluorescein–horseradish peroxidase conjugate binds to either a Fluoresceinated peptide in the lysate or a Fluorescein attached to the wells of a microtiter plate in a competitive fashion. After washing, solid-phase peroxidase activity is measured and inversely related to the amount of FITC-labeled peptide present. To demonstrate the assay, the interaction of a FITC-labeled bombesin-like peptide with the gastrin-releasing peptide receptor on PC-3 and HT-29 cells was investigated. Using PC-3 cells, we obtained similar displacement curves and numbers of binding sites per cell by both the FITC–hapten immunoassay and a reference radioreceptor assay. The FITC–hapten immunoassay is a sensitive and versatile method, since the same commercially available reagents can be used to assess interactions between any peptide and any receptor. In addition, the FITC-labeled peptide can be used to visualize receptors in fluorescent-activated cell sorting or fluorescent microscopy.

  • Fluorescein Isothiocyanate hapten immunoassay for determination of peptide cell interactions
    Analytical Biochemistry, 2004
    Co-Authors: Kimberly A. Kelly, Fred Reynolds, Ralph Weissleder, Lee Josephson
    Abstract:

    Abstract We have developed a Fluorescein Isothiocyanate (FITC)–hapten immunoassay, where a FITC-labeled peptide binding to a cell is assayed as the amount of immunoreactive Fluorescein present in a cell lysate. An antiFluorescein–horseradish peroxidase conjugate binds to either a Fluoresceinated peptide in the lysate or a Fluorescein attached to the wells of a microtiter plate in a competitive fashion. After washing, solid-phase peroxidase activity is measured and inversely related to the amount of FITC-labeled peptide present. To demonstrate the assay, the interaction of a FITC-labeled bombesin-like peptide with the gastrin-releasing peptide receptor on PC-3 and HT-29 cells was investigated. Using PC-3 cells, we obtained similar displacement curves and numbers of binding sites per cell by both the FITC–hapten immunoassay and a reference radioreceptor assay. The FITC–hapten immunoassay is a sensitive and versatile method, since the same commercially available reagents can be used to assess interactions between any peptide and any receptor. In addition, the FITC-labeled peptide can be used to visualize receptors in fluorescent-activated cell sorting or fluorescent microscopy.

Lan Luan - One of the best experts on this subject based on the ideXlab platform.