Abstract: Methods, reagents, and kits are provided that permit flow cytometric determination of the phosphorylation status of retinoblastoma susceptibility gene protein (pRB) in individual cells. Methods are described that permit the hypophosphorylated, active, form of pRB to be measured either as an absolute quantity or as a proportion of total cellular pRB. Further described are methods that permit pRB phosphorylation status to be correlated with cell cycle phase and with protein components of the cell cycle. Screening of chemical compounds for antiproliferative and antineoplastic activity using the flow cytometric assays is demonstrated. Reagent kits that facilitate the subject methods are also provided.
Type:
Grant
Filed:
September 29, 2004
Date of Patent:
July 4, 2006
Assignee:
Becton Dickinson and Company
Inventors:
Zbigniew Darzynkiewicz, Frank Traganos, Gloria Juan, Stefan Gruenwald
Abstract: Methods, reagents, and kits are provided that permit flow cytometric determination of the phosphorylation status of retinoblastoma susceptibility gene protein (pRB) in individual cells. Methods are described that permit the hypophosphorylated, active, form of pRB to be measured either as an absolute quantity or as a proportion of total cellular pRB. Further described are methods that permit pRB phosphorylation status to be correlated with cell cycle phase and with protein components of the cell cycle. Screening of chemical compounds for antiproliferative and antineoplastic activity using the flow cytometric assays is demonstrated. Reagent kits that facilitate the subject methods are also provided.
Type:
Grant
Filed:
February 24, 1999
Date of Patent:
November 23, 2004
Assignee:
Becton, Dickinson and Company
Inventors:
Zbigniew Darzynkiewicz, Frank Traganos, Gloria Juan, Stefan Gruenwald