@article{872c8dd2873e4ebd8736533cfee03228,
title = "Quantitation of cytotoxic rat antibodies by post-assay labeling of mitogen-induced target blasts with [3H]thymidine",
abstract = "Cytotoxic rat alloantibodies were quantitated using concanavalin-A induced blasts as target cells. [3H]thymidine incorporation by such cells was linearly related to their number. Serial dilutions of cytotoxic antisera were incubated with a small number of target cells in presence of rabbit, guinea pig or rat complement. Following a short incubation, cultures were pulsed with [3H]thymidine to estimate the number of live cells. Cytotoxicity titers were calculated according to conventional von Krogh analysis as the reciprocal of the dilution yielding 50% lysis. Such titers were virtually identical to titers obtained in assays in which the extent of cytolysis was determined by trypan blue or ethidium bromide exclusion. The assay, which is carried out in microtiter plates, is quantitative, economical, and objective. Furthermore, automatic harvesting of the cultures allows the rapid processing of large numbers of samples.",
keywords = "cytotoxicity, post-assay labeling, rat alloantibodies",
author = "Martinelli, {G. P.} and D. Blumenthal and H. Schanzer",
note = "Funding Information: Complement dependent antibody mediated cytotoxicity can be assayed by techniques employing various vital dyes, and fluorescent or radioactive markers to determine the proportion of dead and/or live cells. In dye-exclusion assays, cytolysis is assessed by taking advantage of the fact that dead cells do not exclude dyes such as trypan blue, eosin or ethidium bromide (Amos et al., 1969; Edidin, 1970; Terasaki et al., 1978). In dye-inclusion assays, live cells can be scored by their ability to retain neutral red (Filman et al., 1975), or to accumulate fluorescein when exposed to fluorescein-diacetate (fluorochromasia) (Rotman and Papermaster, 1966; Celada and Rotman, 1967). The phenomenon of fluorochromasia has also been used in dye-release tests in which target cells are labeled with fluorescein-diacetate prior to the assay. Upon plasma membrane damage, the accumulated fluorescein is 1 This work was supported in part by a grant from the Jack Martin Fund. 2 Supported by Training Grant no. T35 M 07420 from the National Institute of Health.",
year = "1984",
month = feb,
day = "24",
doi = "10.1016/0022-1759(84)90084-X",
language = "English",
volume = "67",
pages = "53--61",
journal = "Journal of Immunological Methods",
issn = "0022-1759",
publisher = "Elsevier B.V.",
number = "1",
}