PMC:17821 / 25784-26938 JSONTXT

{"project":"2_test","denotations":[{"id":"11056679-8594005-4381694","span":{"begin":147,"end":149},"obj":"8594005"}],"text":"Measurement of IL-1α and IL-1β protein levels\nIL-1 culture supernatants were measured in duplicate by a nonequilibrium RIA as described elsewhere [41]. Briefly, 100 μl polyclonal rabbit anti-murine IL-1α and IL-1β (diluted in RIA buffer, pH 7.4) was added to 100 μl of samples and standards and kept on ice. After vortexing, the tubes were incubated at 4°C. After 24 h, 100 μl of the appropriate 125I-labelled IL-1α and β containing approximately 10 000 cpm was added to each tube, and incubation was continued for a further 24 h at 4°C. RIA buffer (750 μl) containing 9% (w/v) polyethylene glycol 6000 (Merck Diagnostica, Darmstadt, Germany) and 3% (w/v) goat anti-rabbit serum was added, to separate bound and free tracer. The tubes were incubated for 20 min at room temperature. After centrifugation at 1500 × \t\t\t\t\t\tg \t\t\t\t\t for 15 min, supernatants were quickly drained on adsorbent paper. A gamma-counter was used to count the radioactivity remaining on the paper. The radioactivity in control tubes (the nonspecific binding activity) was subtracted from samples and standards. The detection limit of the assay was 20 pg/ml for both IL-1α and IL-1β."}

{"project":"Colil","denotations":[{"id":"T45","span":{"begin":147,"end":149},"obj":"8594005"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/docs/sourcedb/PubMed/sourceid/"}],"text":"Measurement of IL-1α and IL-1β protein levels\nIL-1 culture supernatants were measured in duplicate by a nonequilibrium RIA as described elsewhere [41]. Briefly, 100 μl polyclonal rabbit anti-murine IL-1α and IL-1β (diluted in RIA buffer, pH 7.4) was added to 100 μl of samples and standards and kept on ice. After vortexing, the tubes were incubated at 4°C. After 24 h, 100 μl of the appropriate 125I-labelled IL-1α and β containing approximately 10 000 cpm was added to each tube, and incubation was continued for a further 24 h at 4°C. RIA buffer (750 μl) containing 9% (w/v) polyethylene glycol 6000 (Merck Diagnostica, Darmstadt, Germany) and 3% (w/v) goat anti-rabbit serum was added, to separate bound and free tracer. The tubes were incubated for 20 min at room temperature. After centrifugation at 1500 × \t\t\t\t\t\tg \t\t\t\t\t for 15 min, supernatants were quickly drained on adsorbent paper. A gamma-counter was used to count the radioactivity remaining on the paper. The radioactivity in control tubes (the nonspecific binding activity) was subtracted from samples and standards. The detection limit of the assay was 20 pg/ml for both IL-1α and IL-1β."}