Figure 2
From: Circulating monocytes from healthy individuals and COPD patients
Monitoring of NFκB activation in monocytes from patients with COPD and controls by ELISA-based TransAM kitAMCell nuclear extracts were prepared from monocytes isolated from COPD patients and controls with and without PiZZ AAT deficiency. NFκB activity assay was performed in a 96-well plates with 10 μg of cell extract per well. Absorbance was measured by spectrophotometry at 405 nm. A positive control performed by using a nuclear extract derived from Jurkat cells. This extract is optimized to give a strong signal when used at 2.5 μg/well. Specificity controls were performed by adding a molar excess (20 pmol/well] of mutant NFκB oligonucleotide [the positive signal remained unaffected] and wild-type NFκB oligonucleotide (a signal was abolished). The bars represent mean values of three independent measurements ± SD.