An improved in vitro assay for lymphotoxin |
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| Authors: | J J Kramer G A Granger |
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| Affiliation: | 1. Inmunology Unit, Complejo Asistencial Universitario e Instituto de Biomedicina Universidad de León (IBIOMED), León, Spain;2. Department of Dermatology, Hospital de la Santa Creu i Sant Pau, Barcelona, Spain;3. Universitat Autònoma de Barcelona Medical School, Spain;4. Dermatology Department, Instituto de Investigación Sanitaria La Princesa (IIS-IP), Hospital Universitario de la Princesa, Madrid, Spain;5. Arthritis Unit, Rheumatology Dpt.,Hospital Clinic and IDIBAPS, Barcelona, Spain;6. Rheumatology Department, Hospital 12 de Octubre, Instituto de Investigación Hospital 12 de Octubre, Madrid, Spain;7. Gastroenterology Department, La Paz Universitary Hospital, Madrid, Spain;8. Cardiology Department, Hospital Universitario Lucus Augusti, c/Ulises Romero n°1. 27003, Lugo, Spain;9. Hospital Clinic, Institut Clínic d´Oftalmología (ICOF), Barcelona, Spain;10. University of Barcelona, Barcelona, Spain;11. Department of Neurology-Neuroimmunology, Centre d''Esclerosi Múltiple de Catalunya, (Cemcat), Vall d''Hebron Institut de Recerca, Hospital Universitari Vall d''Hebron, Barcelona, Spain;12. Division of Neurology, St. Michael''s Hospital, University of Toronto, Toronto, Canada;13. Crohn''s and colitis Attention Unit, Digestive System Research Unit, Hospital Vall d''Hebron, Barcelona, Spain;14. Hematology Department, Hospital Universitario Vall d''Hebron, Vall d''Hebron Institute of Oncology (VHIO), Barcelona, Spain;15. Oncología Médica, IOB, Complejo Ruber Juan Bravo, Ruber Internacional, Madrid, Spain;p. Infectious Diseases Department, Hospital Clinic-IDIBAPS, Universitat de Barcelona, Barcelona, Spain;q. Novartis Pharma, Basel, Switzerland;r. Novartis pharmaceuticals Spain, Barcelona, Spain;s. Rheumatology División, IDIVAL, Hospital Universitario Marqués de Valdecilla, Santander, Spain;t. University of Cantabria, Santander, Spain |
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| Abstract: | An In Vitro Microassay for Lymphotoxin (LT) is described. Target cell monolayers are established in the wells of a microtiter assay plate at a density of 100–500/well. Dilutions of Lymphotoxin containing medium are placed upon the target cell monolayers. The plates are sealed with a gas impervious film, and the cells are incubated for 24–48 hr. The cell numbers in each well are established initially, and after the incubation period. The percent destruction is based upon total cell counts of experimental wells compared to control wells. Comparison of the assay to the previously described assay, which is based upon ability of cells to incorporate 14C amino acids into protein, with several batches of LT, shows the microassay to be 25 × more sensitive than the latter. Other advantages and disadvantages of the assay system are also described. |
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