PMC:17821 / 24428-25782 JSONTXT

{"project":"2_test","denotations":[{"id":"11056679-3494788-4381693","span":{"begin":111,"end":113},"obj":"3494788"}],"text":"Bioassay for IL-1\nIL-1 activity was measured in the one-stage bioassay for IL-1 as described by Gearing et al [40]. The assay was performed as a culture of the IL-1-specific thymoma cell EL-4, designated NOB-1, which produces IL-2 and IL-4 in response to IL-1, in co-culture with the lymphokine-responder CTLL line. Briefly, EL-4 cells were washed twice and resuspended at 2.5 × 105 cells per ml RPMI medium containing 5% FCS. The cells were distributed into 96-well microtitre plates at 2.5 × 104 cells per well in 100-μl volumes. CTLL cells (4 × 103) suspended in 50 μl RPMI medium were added, followed by appropriate dilutions of test sample to a final volume of 200 μl. After 20 h, 0.5 μCi 3H-thymidine (specific activity 20 Ci/mmol; Amersham Nederland BV, 's Hertogenbosch, The Netherlands) was added to each well. After a 3-h incubation, the contents were harvested, and incorporated activity was determined. The EL-4 line, from which NOB-1 was derived, does not incorporate thymidine, because it is deficient in thymidine kinase, and therefore the incorporation of 3H-thymidine is a measure of only CTLL proliferation. Maximal 3H-thymidine incorporation in the bioassay in the presence of IL-1 was between 15 000 and 20 000 cpm. Culture media contained only minor concentrations of IL-2 or IL-4, as was assessed by testing samples with CTLL alone."}

{"project":"Colil","denotations":[{"id":"T44","span":{"begin":111,"end":113},"obj":"3494788"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/docs/sourcedb/PubMed/sourceid/"}],"text":"Bioassay for IL-1\nIL-1 activity was measured in the one-stage bioassay for IL-1 as described by Gearing et al [40]. The assay was performed as a culture of the IL-1-specific thymoma cell EL-4, designated NOB-1, which produces IL-2 and IL-4 in response to IL-1, in co-culture with the lymphokine-responder CTLL line. Briefly, EL-4 cells were washed twice and resuspended at 2.5 × 105 cells per ml RPMI medium containing 5% FCS. The cells were distributed into 96-well microtitre plates at 2.5 × 104 cells per well in 100-μl volumes. CTLL cells (4 × 103) suspended in 50 μl RPMI medium were added, followed by appropriate dilutions of test sample to a final volume of 200 μl. After 20 h, 0.5 μCi 3H-thymidine (specific activity 20 Ci/mmol; Amersham Nederland BV, 's Hertogenbosch, The Netherlands) was added to each well. After a 3-h incubation, the contents were harvested, and incorporated activity was determined. The EL-4 line, from which NOB-1 was derived, does not incorporate thymidine, because it is deficient in thymidine kinase, and therefore the incorporation of 3H-thymidine is a measure of only CTLL proliferation. Maximal 3H-thymidine incorporation in the bioassay in the presence of IL-1 was between 15 000 and 20 000 cpm. Culture media contained only minor concentrations of IL-2 or IL-4, as was assessed by testing samples with CTLL alone."}