Tissue factor activity in HeLa cells measured with a continuous chromogenic assay and ELISA reader.
Tissue factor activity expressed by Hela cells cultured in 96-well plates has been quantitated in situ using a continuous spectrophotometric assay. Following the assay, cells assayed without physical disruption remained as viable as cells not subjected to the assay. Very little (or no) tissue factor was expressed in nondisrupted cells relative to that available in cells disrupted by freeze-thawing and sonication. Total tissue factor activity (that available in disrupted cells) decreased not as a simple function of time after subculturing, but was inversely related to cell density.
D000818:Animals; D002417:Cattle; D003584:Cytological Techniques; D004797:Enzyme-Linked Immunosorbent Assay; D005167:Factor VII / Q000502:physiology; D015942:Factor VIIa; D005170:Factor X / Q000502:physiology; D006367:HeLa Cells / Q000378:metabolism*; D006801:Humans; D008722:Methods; D008954:Models, Biological; D013053:Spectrophotometry; D013925:Thromboplastin / Q000378:metabolism*; D013997:Time Factors
D005167:Factor VII; D005170:Factor X; D013925:Thromboplastin; D015942:Factor VIIa