Mouse anti human il 1β antibody

Two HuProt human protein microarray slides (v.2.0) containing 19,951 probe sets spotted in duplicate were purchased from CDI Laboratories (Mayaguez, PR). Microarray slides were preincubated in block buffer (2% BSA and 0.1% Tween in PBS) for 2 h at room temperature. Slides were then aspirated and incubated with recombinant human pro-IL-1β (10 μg/ml) diluted in reagent diluent (PBS containing Ca²⁺ with 0.1% Tween) or reagent diluent alone (negative control) for 1 h at room temperature. Both slides were aspirated and washed three times with reagent diluent. The microarray slides were then incubated with mouse anti-human IL-1β antibody (R&D Systems) diluted in reagent diluent. As before, slides were aspirated and washed three times. Finally, slides were incubated with an Alexa Fluor 633-conjugated goat anti-mouse secondary antibody (2 μg/ml) in reagent diluent for 1 h at room temperature, followed by a final three washes. The microarrays were spun dry at 1500 rpm for 3 min and scanned using a GenePix 4000B microarray scanner (Molecular Devices, Sunnyvale, CA).